JP7126269B2 - Methods for treating cancer comprising TIGIT binding agents - Google Patents

Description

ATCC PTA-122346 ATCC PTA-122347

FIELD OF THE INVENTION The present invention relates generally to methods for treating cancer using agents that bind to human TIGIT, and in particular, using antibodies that specifically bind to the extracellular domain of TIGIT.

BACKGROUND OF THE INVENTION The basis of immunotherapy is the manipulation and/or modulation of the immune system, including both innate and adaptive immune responses. The general purpose of immunotherapy is to treat disease by controlling the immune response to "foreign agents," eg pathogens or tumor cells. In some cases, however, immunotherapy is used to treat autoimmune diseases that can result from aberrant immune responses against proteins, molecules, and/or tissues normally present in the body. Immunotherapy may involve agents and methods for inducing or enhancing a specific immune response, or may involve methods for inhibiting or reducing a specific immune response.

The immune system is a highly complex system composed of numerous cell types including, but not limited to, T cells, B cells, natural killer cells, antigen presenting cells, dendritic cells, monocytes, and macrophages. . These cells have complex and sophisticated systems to control these interactions and responses. These cells employ activating and inhibitory mechanisms and feedback loops to stem the response and prevent the negative consequences of an uncontrolled immune response (eg, autoimmune disease) from being allowed.

The concept of cancer immunosurveillance is based on the theory that the immune system can recognize tumor cells, mount an immune response, and suppress tumor development and/or progression. However, it is clear that many cancer/tumor cells have developed immune system evasion mechanisms that may allow unsuppressed growth of these cells. Cancer/tumor immunotherapies are novel and capable of activating and/or boosting the immune system to achieve a more effective attack on tumor cells and boost tumor cell killing and/or inhibition of tumor growth. It focuses on developing novel agents.

BRIEF SUMMARY OF THE INVENTION The present invention includes, but is not limited to, antibodies that specifically bind to the extracellular domain of a T-cell immunoreceptor with Ig and ITIM domains (TIGIT) using agents that bind to TIGIT. , provides methods for treating cancer (ie, for inhibiting tumor growth). In certain embodiments, the agent is a TIGIT antagonist. In some embodiments, the method comprises using a TIGIT binding agent to induce, activate, promote, increase, enhance, or prolong an immune response against cancer and/or tumors. In some embodiments, the method comprises using a TIGIT binding agent to inhibit tumor growth. In some embodiments, the method comprises using a TIGIT binding agent to treat cancer. In some embodiments, the method comprises using the TIGIT binding agent in combination with at least one additional therapeutic agent.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT binding agent, wherein the tumor is colorectal cancer (CRC), e.g. Frequency microsatellite instability-high colorectal cancer (MSI CRC) or microsatellite stable colorectal cancer (MSS CRC), triple-negative breast cancer (TNBC), Merkel cell carcinoma, uterus Endometrial cancer or esophageal cancer. In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, wherein the tumor comprises frequent microsatellites An antibody that is unstable colorectal cancer, microsatellite-stable colorectal cancer, triple-negative breast cancer, Merkel cell carcinoma, endometrial cancer, or esophageal cancer and that binds to human TIGIT is TSDYAWN (SEQ ID heavy chain CDR1 containing NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

, light chain CDR2 with SASYRYT (SEQ ID NO:8), and light chain CDR3 with QQHYSTP (SEQ ID NO:9).

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、高頻度MSI固形腫瘍は、MSI CRC、MSI胃がん、MSI子宮内膜がん、MSI卵巣がん、MSI肝胆道がん、MSI尿路がん、MSI脳がん、またはMSI皮膚がんからなる群より選択される。 In some aspects of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT binding agent, wherein the tumor is a high frequency microsatellite instability (MSI) solid tumor. is. In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, and the antibody that binds to human TIGIT is the heavy chain CDR1 containing TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the common MSI solid tumor is MSI CRC, MSI gastric cancer, MSI endometrial cancer, MSI ovarian cancer, MSI hepatobiliary cancer, MSI urinary tract cancer, MSI brain cancer, or MSI selected from the group consisting of skin cancer;

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、併用療法は抗PD-1抗体を含む。一部の態様において、併用療法は抗PD-L1抗体を含む。 In some embodiments of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist, wherein the tumor is , high-frequency microsatellite-unstable colorectal cancer, microsatellite-stable colorectal cancer, triple-negative breast cancer, Merkel cell carcinoma, endometrial cancer, or esophageal cancer. In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or a PD-L1 antagonist. wherein the tumor is colorectal cancer (CRC), e.g., high frequency microsatellite unstable colorectal cancer or microsatellite stable colorectal cancer, triple negative breast cancer, Merkel cell carcinoma, endometrial An antibody that is cancer or esophageal cancer and that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, combination therapy comprises an anti-PD-1 antibody. In some embodiments, combination therapy comprises an anti-PD-L1 antibody.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、併用療法は抗PD-1抗体を含む。一部の態様において、併用療法は抗PD-L1抗体を含む。一部の態様において、高頻度MSI固形腫瘍は、MSI CRC、MSI胃がん、MSI子宮内膜がん、MSI卵巣がん、MSI肝胆道がん、MSI尿路がん、MSI脳がん、またはMSI皮膚がんからなる群より選択される。 In some embodiments of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist, wherein the tumor is It is a high frequency MSI solid tumor. In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or a PD-L1 antagonist. wherein the tumor is a high frequency microsatellite unstable solid tumor and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, combination therapy comprises an anti-PD-1 antibody. In some embodiments, combination therapy comprises an anti-PD-L1 antibody. In some embodiments, the common MSI solid tumor is MSI CRC, MSI gastric cancer, MSI endometrial cancer, MSI ovarian cancer, MSI hepatobiliary cancer, MSI urinary tract cancer, MSI brain cancer, or MSI selected from the group consisting of skin cancer;

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、腫瘍は、PD-1アンタゴニストまたはPD-L1アンタゴニストを用いる処置に対して抵抗性または不応性であった。本明細書で用いられるように、腫瘍は、作用物質を用いる処置に対して抵抗性または不応性であり、作用物質を用いる処置の最中または後に腫瘍成長進行がある。一部の態様において、腫瘍は、単一の作用物質として投与される作用物質を用いる処置に対して抵抗性または不応性である。一部の態様において、腫瘍は、第一選択治療薬または第二選択治療薬として投与される作用物質を用いる処置に対して抵抗性または不応性である。一部の態様において、腫瘍は、(例えば、単一の作用物質として)抗PD-1抗体を用いる処置に対して抵抗性または不応性である。一部の態様において、腫瘍は、(例えば、単一の作用物質として)抗PD-L1抗体を用いる処置に対して抵抗性または不応性である。一部の態様において、PD-1アンタゴニストまたはPD-L1アンタゴニストを用いる処置に対して抵抗性または不応性の腫瘍は、黒色腫、非小細胞肺がん(NSCLC)、腎細胞がん、頭頸部扁平上皮がん、尿路上皮がん、結腸直腸がん(例えば、MSIまたはDNAミスマッチ修復欠損(dMMR)転移性CRC)、および肝細胞がんからなる群より選択される。 In some embodiments of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT-binding agent, wherein the tumor employs a PD-1 antagonist or a PD-L1 antagonist. Resistant or refractory to treatment. In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, wherein the tumor is a PD-1 antagonist or an antibody that is resistant or refractory to treatment with a PD-L1 antagonist and that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the tumor was resistant or refractory to treatment with a PD-1 antagonist or PD-L1 antagonist. As used herein, a tumor is resistant or refractory to treatment with an agent and has tumor growth progression during or after treatment with an agent. In some embodiments, the tumor is resistant or refractory to treatment with agents administered as a single agent. In some embodiments, the tumor is resistant or refractory to treatment with an agent administered as a first-line or second-line therapeutic. In some embodiments, the tumor is resistant or refractory to treatment with an anti-PD-1 antibody (eg, as a single agent). In some embodiments, the tumor is resistant or refractory to treatment with an anti-PD-L1 antibody (eg, as a single agent). In some embodiments, the tumor resistant or refractory to treatment with a PD-1 antagonist or PD-L1 antagonist is melanoma, non-small cell lung cancer (NSCLC), renal cell carcinoma, head and neck squamous cell selected from the group consisting of cancer, urothelial carcinoma, colorectal cancer (eg MSI or DNA mismatch repair deficient (dMMR) metastatic CRC), and hepatocellular carcinoma.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、腫瘍は、第一選択治療薬または第二選択治療薬として投与された作用物質に対して抵抗性または不応性である。一部の態様において、腫瘍は、単一の作用物質としてPD-1アンタゴニストを用いる処置に対して抵抗性または不応性であるが、TIGIT結合物質と組み合わせてこの作用物質を用いる処置に応答性である。一部の態様において、腫瘍は、単一の作用としてPD-L1アンタゴニストを用いる処置に対して抵抗性または不応性であるが、TIGIT結合物質と組み合わせてこの作用物質を用いる処置に応答性である。一部の態様において、腫瘍は、(例えば、単一の作用物質として)抗PD-1抗体を用いる処置に対して抵抗性または不応性である。一部の態様において、腫瘍は、(例えば、単一の作用物質として)抗PD-L1抗体を用いる処置に対して抵抗性または不応性である。一部の態様において、併用療法は抗PD-1抗体を含む。一部の態様において、併用療法は抗PD-L1抗体を含む。一部の態様において、PD-1アンタゴニストまたはPD-L1アンタゴニストを用いる処置に対して抵抗性または不応性の腫瘍は、黒色腫、NSCLC、腎細胞がん、頭頸部扁平上皮がん、尿路上皮がん、結腸直腸がん(例えば、MSIまたはdMMR転移性CRC)、および肝細胞がんからなる群より選択される。 In some embodiments of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist, wherein the tumor is , resistant or refractory to treatment with PD-1 antagonists or PD-L1 antagonists as single agents. In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or a PD-L1 antagonist. wherein the tumor is resistant or refractory to treatment with a PD-1 antagonist or PD-L1 antagonist as a single agent, and the antibody that binds to human TIGIT is TSDYAWN (SEQ ID heavy chain CDR1 containing NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the tumor is resistant or refractory to agents administered as first-line or second-line therapeutic agents. In some embodiments, the tumor is resistant or refractory to treatment with a PD-1 antagonist as a single agent, but responsive to treatment with this agent in combination with a TIGIT binding agent. be. In some embodiments, the tumor is resistant or refractory to treatment with a PD-L1 antagonist as a single action, but responsive to treatment with this agent in combination with a TIGIT binding agent. . In some embodiments, the tumor is resistant or refractory to treatment with an anti-PD-1 antibody (eg, as a single agent). In some embodiments, the tumor is resistant or refractory to treatment with an anti-PD-L1 antibody (eg, as a single agent). In some embodiments, combination therapy comprises an anti-PD-1 antibody. In some embodiments, combination therapy comprises an anti-PD-L1 antibody. In some embodiments, the tumor resistant or refractory to treatment with a PD-1 antagonist or PD-L1 antagonist is melanoma, NSCLC, renal cell carcinoma, head and neck squamous cell carcinoma, urothelium selected from the group consisting of cancer, colorectal cancer (eg MSI or dMMR metastatic CRC), and hepatocellular carcinoma.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、対象は、PD-1アンタゴニストまたはPD-L1アンタゴニストを用いて以前に処置されたことがあり、処置中にまたは処置後に疾患進行があった。一部の態様において、PD-1アンタゴニストまたはPD-L1アンタゴニストを用いる処置の後に腫瘍成長再発がある。本明細書で使用する場合、腫瘍成長の再発とは、処置後でかつがんが検出されない期間の後にがんが再び現れることと定義される。がんは、最初に発見された場所で再発してもよく、体内のどこか他の場所で発見されてもよい。一部の態様において、再発は局所再発、局部再発、または遠隔再発である。一部の態様において、対象は抗PD-1抗体を用いて以前に処置されたことがある。一部の態様において、対象は抗PD-L1抗体を用いて以前に処置されたことがある。一部の態様において、腫瘍は、黒色腫、NSCLC、腎細胞がん、頭頸部扁平上皮がん、尿路上皮がん、結腸直腸がん(例えば、MSIまたはdMMR転移性CRC)、および肝細胞がんからなる群より選択される。 In some embodiments of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT binding agent, wherein the subject is a PD-1 antagonist or Has been previously treated with a PD-L1 antagonist. In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, wherein the subject has a single effect Antibodies that have been previously treated with a PD-1 antagonist or a PD-L1 antagonist as agents and that bind to human TIGIT have a heavy chain CDR1 containing TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the subject has been previously treated with a PD-1 antagonist or PD-L1 antagonist and has had disease progression during or after treatment. In some embodiments, there is tumor growth recurrence following treatment with a PD-1 antagonist or PD-L1 antagonist. As used herein, recurrence of tumor growth is defined as the reappearance of cancer after treatment and after a period of no cancer detection. Cancer may recur where it was first found, or it may be found elsewhere in the body. In some embodiments, the recurrence is local recurrence, local recurrence, or distant recurrence. In some embodiments, the subject has been previously treated with an anti-PD-1 antibody. In some embodiments, the subject has been previously treated with an anti-PD-L1 antibody. In some embodiments, the tumor is melanoma, NSCLC, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial carcinoma, colorectal cancer (eg, MSI or dMMR metastatic CRC), and hepatocellular carcinoma. selected from the group consisting of cancer;

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、対象は、PD-1アンタゴニストまたはPD-L1アンタゴニストを用いて以前に処置されたことがあり、処置中にまたは処置後に疾患進行があった。一部の態様において、PD-1アンタゴニストまたはPD-L1アンタゴニストを用いる処置の後に腫瘍成長再発がある。一部の態様において、再発は局所再発、局部再発、または遠隔再発である。一部の態様において、併用療法は、PD-1アンタゴニストまたはPD-L1アンタゴニストを用いる処置に対して腫瘍を「再増感させる(resensitize)」。一部の態様において、対象は抗PD-1抗体を用いて以前に処置されたことがある。一部の態様において、対象は抗PD-L1抗体を用いて以前に処置されたことがある。一部の態様において、併用療法は抗PD-1抗体を含む。一部の態様において、併用療法は抗PD-L1抗体を含む。一部の態様において、腫瘍は、黒色腫、NSCLC、腎細胞がん、頭頸部扁平上皮がん、尿路上皮がん、結腸直腸がん(例えば、MSIまたはdMMR転移性CRC)、および肝細胞がんからなる群より選択される。 In some embodiments of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist, wherein the subject is , has been previously treated with a PD-1 antagonist or a PD-L1 antagonist as a single agent. In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or a PD-L1 antagonist. wherein the subject has been previously treated with a PD-1 antagonist or a PD-L1 antagonist as a single agent, and the antibody that binds human TIGIT is TSDYAWN (SEQ ID NO: 4) heavy chain CDR1, including

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the subject has been previously treated with a PD-1 antagonist or PD-L1 antagonist and has had disease progression during or after treatment. In some embodiments, there is tumor growth recurrence following treatment with a PD-1 antagonist or PD-L1 antagonist. In some embodiments, the recurrence is local recurrence, local recurrence, or distant recurrence. In some embodiments, the combination therapy "resensitizes" the tumor to treatment with a PD-1 antagonist or PD-L1 antagonist. In some embodiments, the subject has been previously treated with an anti-PD-1 antibody. In some embodiments, the subject has been previously treated with an anti-PD-L1 antibody. In some embodiments, combination therapy comprises an anti-PD-1 antibody. In some embodiments, combination therapy comprises an anti-PD-L1 antibody. In some embodiments, the tumor is melanoma, NSCLC, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial carcinoma, colorectal cancer (eg, MSI or dMMR metastatic CRC), and hepatocellular carcinoma. selected from the group consisting of cancer;

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT binding agent, wherein the tumor is poliovirus receptor (PVR) and/or poliovirus. Express receptor-related 2 (PVRL2). In some embodiments, the method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, wherein the cancer is PVR and/or An antibody that expresses PVRL2 and binds to human TIGIT is a heavy chain CDR1 containing TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、腫瘍は、黒色腫、NSCLC、腎細胞がん、頭頸部扁平上皮がん、尿路上皮がん、結腸直腸がん(例えば、MSIまたはdMMR)転移性CRC、および肝細胞がんからなる群より選択される。 In some embodiments of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or a PD-L1 antagonist, the cancer express PVR and/or PVRL2. In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or a PD-L1 antagonist. wherein the cancer expresses PVR and/or PVRL2 and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the tumor is melanoma, NSCLC, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial carcinoma, colorectal cancer (eg, MSI or dMMR) metastatic CRC, and hepatocellular carcinoma. selected from the group consisting of cancer;

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments of the invention, a method of inhibiting tumor growth in a subject comprises administering a therapeutically effective amount of a TIGIT binding agent to the subject, wherein the tumor comprises tumor infiltrating lymphocytes (TILs). In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, wherein the tumor is free of tumor-infiltrating lymphocytes. The antibody comprising and binding to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist, wherein the tumor is Contains tumor-infiltrating lymphocytes. In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or a PD-L1 antagonist. wherein the tumor comprises tumor infiltrating lymphocytes (TILs), and the antibody that binds to human TIGIT comprises a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some aspects of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT binding agent, wherein the tumor comprises regulatory T cells (Treg). In some embodiments, the method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, the tumor comprises Tregs, and Antibodies that bind to human TIGIT are heavy chain CDR1 containing TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments of the invention, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist, wherein the tumor is Including Tregs. In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or a PD-L1 antagonist. wherein the tumor comprises Tregs and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

In some embodiments of the methods described herein, the tumor is lung tumor, liver tumor, breast tumor, renal cell carcinoma/kidney tumor, prostate tumor, gastrointestinal/gastric tumor, melanoma, neck tumor, It is selected from the group consisting of bladder tumor, glioblastoma, head and neck tumor, pancreatic tumor, ovarian tumor, colorectal tumor, endometrial tumor, anal tumor and esophageal tumor.

In some embodiments, the lung tumor comprises NSCLC, eg, NSCLC squamous cell or NSCLC adenocarcinoma.

In some embodiments, the breast tumor comprises triple negative breast cancer (TNBC).

In some embodiments, the tumor is a high MSI solid tumor. In some embodiments, the high frequency microsatellite unstable solid tumor is MSI colorectal cancer (MSI CRC), MSI gastric cancer, MSI endometrial cancer, MSI ovarian cancer, MSI hepatobiliary cancer, MSI urinary tract cancer cancer, MSI brain cancer, or MSI skin cancer. In some embodiments, the frequent MSI solid tumor is MSI CRC.

In some embodiments, the tumor is a microsatellite stable (MSS) tumor. In some embodiments, the tumor is microsatellite stable colorectal cancer (MSS CRC).

In some embodiments, the tumor is melanoma, NSCLC, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial carcinoma, colorectal cancer (eg, MSI or dMMR metastatic CRC), and hepatocellular carcinoma. selected from the group consisting of cancer;

を含む重鎖CDR2、および

を含む重鎖CDR3、ならびに/または

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments of the methods described herein, the agent is an antibody that specifically binds to the extracellular domain of human TIGIT, and the antibody that specifically binds to the extracellular domain of human TIGIT is TSDYAWN heavy chain CDR1 comprising (SEQ ID NO:4),

heavy chain CDR2 containing, and

a heavy chain CDR3 comprising, and/or

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

In some embodiments of the methods described herein, the agent is an antibody that specifically binds to the extracellular domain of TIGIT, said antibody having at least 90% sequence identity to SEQ ID NO:10 and/or a light chain variable region having at least 90% sequence identity with SEQ ID NO:11. In some embodiments, the antibody has a heavy chain variable region having at least 95% sequence identity with SEQ ID NO:10 and/or a light chain variable region having at least 95% sequence identity with SEQ ID NO:11. including. In some embodiments, the antibody comprises a heavy chain variable region comprising SEQ ID NO:10 and a light chain variable region comprising SEQ ID NO:11.

In some embodiments of the methods described herein, the TIGIT binding agent is a monoclonal antibody, humanized antibody, human antibody, recombinant antibody, chimeric antibody, bispecific antibody, antibody fragment comprising an antigen binding site, An antibody that is an IgG antibody, an IgG1 antibody, an IgG2 antibody, or an IgG4 antibody. In some embodiments, said antibody is monovalent. In some embodiments, said antibody is bivalent. In some embodiments, the antibody is monospecific. In some embodiments, said antibody is bispecific.

In some embodiments of the methods described herein, the antibody that specifically binds human TIGIT comprises the heavy chain amino acid sequence of SEQ ID NO:13 and the light chain amino acid sequence of SEQ ID NO:15. In some embodiments, the antibody that specifically binds human TIGIT comprises the heavy chain amino acid sequence of SEQ ID NO:17 and the light chain amino acid sequence of SEQ ID NO:15.

In some embodiments of the methods described herein, the antibody that specifically binds human TIGIT does not bind mouse TIGIT. In some embodiments, an antibody that specifically binds human TIGIT does not bind rat TIGIT. In some embodiments, an antibody that specifically binds human TIGIT does not bind rabbit TIGIT. In some embodiments, an antibody that specifically binds human TIGIT does not bind marmoset TIGIT. In some embodiments, an antibody that specifically binds human TIGIT does not bind canine TIGIT. In some embodiments, an antibody that specifically binds human TIGIT does not bind porcine TIGIT. In some embodiments, an antibody that specifically binds human TIGIT does not bind cynomolgus monkey TIGIT. In some embodiments, an antibody that specifically binds human TIGIT does not bind rhesus TIGIT.

In some embodiments of the methods described herein, the antibody that specifically binds human TIGIT comprises heavy and light chain variable regions from antibody OMP-313M32. In some embodiments, the antibody comprises a heavy chain variable region encoded by a plasmid deposited with the ATCC under designation number PTA-122346. In some embodiments, the antibody comprises a polypeptide comprising a heavy chain variable region encoded by a plasmid deposited with the ATCC under designation number PTA-122346. In some embodiments, the antibody comprises a light chain variable region encoded by a plasmid deposited with the ATCC under designation number PTA-122347. In some embodiments, the antibody comprises a light chain encoded by a plasmid deposited with the ATCC under designation number PTA-122347. In some embodiments, the antibody comprises a heavy chain variable region encoded by a plasmid deposited with the ATCC under designation number PTA-122346 and a light chain encoded by a plasmid deposited with the ATCC under designation number PTA-122347. Contains variable regions. In some embodiments, the antibody comprises a polypeptide encoded by the plasmid deposited with the ATCC as PTA-122346 and a polypeptide encoded by the plasmid deposited with the ATCC under designation number PTA-122347.

In some embodiments of the methods described herein, the TIGIT binding agent is monovalent. In some embodiments, the TIGIT binding agent is bivalent. In some embodiments, the TIGIT binding agent is monospecific. In some embodiments, the TIGIT binding agent is bispecific. In some embodiments, the bispecific agent is a heterodimeric agent or heterodimeric molecule. In some embodiments, the heterodimeric agent comprises an antibody that specifically binds TIGIT. In some embodiments, the bispecific agent is a homodimeric agent or homodimeric molecule. In some embodiments, the homodimeric agent comprises an antibody that specifically binds TIGIT.

In some embodiments of the methods described herein, the TIGIT binding agent is an antibody that competes with the antibodies described herein for specific binding to human TIGIT. In some embodiments, the TIGIT binding agent binds to the same epitope on human TIGIT as the antibodies described herein. In some embodiments, the TIGIT binding agent binds to an epitope on human TIGIT that overlaps with the epitope on TIGIT that is bound by an antibody described herein. In some embodiments, the TIGIT binding agent binds to an epitope comprising amino acids in SEQ ID NO:27. In some embodiments, the TIGIT binding agent binds to an epitope comprising amino acids in SEQ ID NO:28. In some embodiments, the TIGIT binding agent binds to an epitope comprising amino acids in SEQ ID NO:27 and SEQ ID NO:28. In some embodiments, the TIGIT binding agent binds to an epitope comprising amino acids Q62 and I109 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent binds to an epitope comprising amino acids Q62 and T119 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent binds to an epitope comprising amino acids Q64 and I109 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent binds to an epitope comprising amino acids Q64 and T119 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent binds to an epitope comprising amino acids Q62, Q64, and I109 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent binds to an epitope comprising amino acids Q62, Q64, and T119 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent binds to an epitope comprising amino acids Q62, I109, and T119 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent binds to an epitope comprising amino acids Q64, I109, and T119 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent binds to an epitope comprising amino acids Q62, Q64, I109, and T119 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent is at least one selected from the group consisting of N58, E60, Q62, Q64, L65, F107, I109, H111, T117, T119, G120, and R121 of SEQ ID NO:1. It binds to an epitope containing three amino acids. In some embodiments, the epitope is a conformational epitope. In some embodiments, the TIGIT binding agent binds to an epitope of SEQ ID NO:1 that does not include amino acid V100.

In some embodiments of the methods described herein, the agent that specifically binds to TIGIT is an antibody, and said antibody is part of a bispecific agent. In some embodiments, the bispecific agent comprises a first arm that binds TIGIT and a second arm that binds a second target. In some embodiments, the bispecific agent comprises a first arm that specifically binds TIGIT, and a second arm, the first arm comprising an anti-TIGIT antibody. In some embodiments, the bispecific agent comprises a first arm that binds TIGIT and a second arm that comprises an antigen binding site derived from an antibody. In some embodiments, the bispecific agent has a first arm that binds TIGIT and specific for PD-1, PD-L1, CTLA4, TIM-3, LAG-3, OX-40, or GITR. It includes a second arm that physically connects. In some embodiments, the bispecific agent comprises a first arm that binds TIGIT and a second arm that specifically binds a tumor antigen. In some embodiments, the bispecific agent comprises a first arm that binds TIGIT and a second arm comprising an immune response stimulating agent. In some embodiments, the immune response stimulating agent is granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), granulocyte colony-stimulating factor (G-CSF), interleukin-2. (IL-2), interleukin 3 (IL-3), interleukin 12 (IL-12), interleukin 15 (IL-15), B7-1 (CD80), B7-2 (CD86), 4-1BB ligand, GITRL, OX-40L, anti-CD3 antibody, anti-CTLA4 antibody, anti-PD-1 antibody, anti-PD-L1 antibody, anti-GITR antibody, anti-OX-40 antibody, anti-LAG-3 antibody, and anti-TIM-3 antibody selected from the group consisting of

In some embodiments, the bispecific agent is a heterodimeric agent or heterodimeric molecule. In some embodiments, the bispecific agent is a homodimeric agent or homodimeric molecule. In some embodiments, the heterodimeric molecule comprises a first arm that binds human TIGIT and a second arm that binds a second target. In some embodiments, the heterodimeric molecule comprises a first arm that specifically binds to human TIGIT, and a second arm, the first arm comprising an anti-TIGIT antibody. In some embodiments, the heterodimeric molecule comprises a first arm that binds human TIGIT and a second arm comprising an antigen binding site derived from an antibody that specifically binds a second target. In some embodiments, the heterodimeric molecule is a bispecific antibody. In some embodiments, the heterodimeric molecule comprises a first arm that binds human TIGIT and a second arm that specifically binds a tumor antigen. In some embodiments, the heterodimeric molecule has a first arm that binds human TIGIT and PD-1, PD-L1, CTLA-4, TIM-3, LAG-3, OX-40, 4- 1BB, or contains a second arm that specifically binds to GITR. In some embodiments, the heterodimeric molecule comprises a first arm that binds TIGIT and a second arm that comprises an immunotherapeutic agent. In some embodiments, the immunotherapeutic agent is granulocyte-macrophage colony stimulating factor (GM-CSF), macrophage colony stimulating factor (M-CSF), granulocyte colony stimulating factor (G-CSF), interleukin 2 (IL -2), interleukin 3 (IL-3), interleukin 12 (IL-12), interleukin 15 (IL-15), B7-1 (CD80), B7-2 (CD86), 4-1BB ligand, GITRL, OX-40L, anti-CD3 antibody, anti-CTLA-4 antibody, anti-PD-1 antibody, anti-PD-L1 antibody, anti-4-1BB antibody, anti-GITR antibody, anti-OX-40 antibody, anti-LAG-3 antibody, and anti-TIM-3 antibodies.

In some embodiments of the methods described herein, the agent specifically binds to TIGIT and inhibits binding of TIGIT to PVR. In some embodiments, the agent specifically binds to TIGIT and inhibits or blocks the interaction between TIGIT and PVR. In some embodiments, the agent specifically binds to TIGIT and inhibits binding of TIGIT to PVRL2. In some embodiments, the agent specifically binds to TIGIT and inhibits or blocks the interaction between TIGIT and PVRL2. In some embodiments, the agent specifically binds to TIGIT and inhibits binding of TIGIT to PVRL3. In some embodiments, the agent specifically binds to TIGIT and inhibits or blocks the interaction between TIGIT and PVRL3. In some embodiments, the agent is an antagonist of TIGIT. In some embodiments, the agent specifically binds to TIGIT and inhibits TIGIT signaling. In some embodiments, the agent specifically binds to TIGIT and is an antagonist of signaling through TIGIT. In some embodiments, the agent specifically binds to TIGIT and inhibits TIGIT activation. In some embodiments, the agent specifically binds to TIGIT and inhibits phosphorylation of TIGIT. In some embodiments, the agent specifically binds to TIGIT and reduces cell surface expression of TIGIT.

In some embodiments of the methods described herein, the agent specifically binds to TIGIT and induces, activates, promotes, augments, enhances an immune response, and/or extend. In some embodiments, the immune response is directed against (eg, killing) the tumor or tumor cells. In some embodiments, the agent increases cell-mediated immunity. In some embodiments, the agent increases T cell activity. In some embodiments, the agent increases cytolytic T cell (CTL) activity. In some embodiments, the agent increases natural killer (NK) cell activity. In some embodiments, the agent increases IL-2 production and/or the number of IL-2 producing cells. In some embodiments, the agent increases IFN-γ production and/or the number of IFN-γ-producing cells. In some embodiments, the agent increases a Th1-type immune response. In some embodiments, the agent reduces IL-4 production and/or the number of IL-4 producing cells. In some embodiments, the agent reduces the number of IL-10 and/or IL-10 producing cells. In some embodiments, the agent reduces IL-6 production and/or the number of IL-6 producing cells. In some embodiments, the agent reduces IL-5 production and/or the number of IL-5 producing cells. In some embodiments, the agent reduces Th2-type immune responses. In some embodiments, the agent reduces the number of Treg cells. In some embodiments, the agent decreases Treg activity. In some embodiments, the agent inhibits and/or reduces the suppressive activity of Tregs. In some embodiments, the agent reduces the number of myeloid-derived suppressor cells (MDSCs). In some embodiments, the agent inhibits and/or reduces suppressive activity of MDSCs.

In some embodiments of the methods described herein, the agent specifically binds to TIGIT and inhibits tumor growth. In some embodiments, the agent reduces tumor growth. In some embodiments, the agent reduces tumor growth to an undetectable size. In some embodiments, the agent induces long-term anti-tumor immunity.

In another aspect, the invention provides compositions comprising a TIGIT binding agent for use in the methods described herein. In some aspects, the invention provides pharmaceutical compositions comprising a TIGIT binding agent for use in the methods described herein and a pharmaceutically acceptable carrier.

In certain embodiments of the methods described herein, said TIGIT binding agent is isolated. In certain embodiments, the TIGIT binding agent is substantially pure.

The invention also provides polynucleotides, including polynucleotides encoding TIGIT binding agents. In some embodiments, the polynucleotide is isolated. In some aspects, the invention provides vectors comprising said polynucleotides, and cells comprising said vectors and/or said polynucleotides. In some embodiments, the invention also provides cells comprising or producing a TIGIT binding agent. In some embodiments, the cells are monoclonal cell lines.

In some embodiments of the methods described herein, the method further comprises administering at least one additional therapeutic agent. In some embodiments, the additional therapeutic agent is a chemotherapeutic agent. In some embodiments, the additional therapeutic agent is an antibody. In some embodiments, the additional therapeutic agent is an anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA4 antibody, anti-LAG-3 antibody, or anti-TIM-3 antibody. In some embodiments, the additional therapeutic agent is an inhibitor of the Notch pathway, Wnt pathway, or RSPO/LGR pathway. In some embodiments, the additional therapeutic agent is an immunotherapeutic agent. As used herein, the phrase "immunotherapeutic agent" is used in its broadest sense and refers to substances that directly or indirectly affect or modulate the immune system. In some embodiments, an immunotherapeutic agent is an agent that directly or indirectly stimulates the immune system by inducing activation or increasing the activity of any immune system component. Since the TIGIT binding agent is considered an immunotherapeutic agent, this additional immunotherapeutic agent may be considered a "second" immunotherapeutic agent. In some embodiments, the second immunotherapeutic agent is GM-CSF, M-CSF, G-CSF, IL-2, IL-3, IL-12, IL-15, B7-1 (CD80), B7 -2 (CD86), 4-1BB ligand, GITRL, OX-40 ligand, anti-CD3 antibody, anti-CTLA-4 antibody, anti-CD28 antibody, anti-PD-1 antibody, anti-PD-L1 antibody, anti-4-1BB antibody, selected from the group consisting of anti-GITR antibody, anti-OX-40 antibody, anti-LAG-3 antibody, and anti-TIM-3 antibody. In some embodiments, the second immunotherapeutic agent is GM-CSF, M-CSF, G-CSF, IL-2, IL-3, IL-12, IL-15, B7-1 (CD80), B7 -2(CD86), 4-1BB ligand, GITRL, OX-40 ligand, or fragments thereof. In some embodiments, the second immunotherapeutic agent is a fusion protein comprising at least one copy of the GITRL extracellular domain, OX40 ligand, or 4-1BB ligand.

In some embodiments of the methods described herein, the subject is human. In some embodiments, the subject is at least partially cleared of a tumor or cancer.

In some embodiments of the methods described herein, the tumor or cancer expresses PD-L1. In some embodiments, the method further comprises determining PD-L1 expression levels in the tumor or cancer. In some embodiments, the step of determining PD-L1 expression level is performed prior to being treated with a TIGIT binding agent or prior to being contacted with a TIGIT binding agent. In some embodiments, a TIGIT binding agent is administered to a subject when the tumor or cancer has high levels of PD-L1 expression.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、方法を含む。 In some embodiments, the invention provides a method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, wherein the tumor but head and neck cancer, esophageal/gastroesophageal cancer, gastric cancer, colorectal cancer, anal cancer, hepatocellular carcinoma/liver cancer, cervical cancer, lung cancer (e.g., NSCLC), melanoma , Merkel cell carcinoma, renal cell/renal cancer, bladder cancer, ovarian cancer, pancreatic cancer, endometrial cancer, and triple-negative breast cancer, known MSI solid tumors (including MSI CRC) or MSS colorectal cancer, the tumor is resistant or refractory to treatment with an anti-PD-1 antibody or an anti-PD-L1 antibody, and an antibody that binds to human TIGIT is TSDYAWN (SEQ ID NO :4) the heavy chain CDR1, including

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

In some embodiments of the methods described herein, the subject has been previously treated with an anti-PD-1 antibody or an anti-PD-L1 antibody. In some embodiments, the antibody that specifically binds to the extracellular domain of human TIGIT is about 0.1 mg/kg, about 0.3 mg/kg, about 1 mg/kg, about 1.5 mg/kg, about 2 mg/kg, about A dose of 2.5 mg/kg, about 3 mg/kg, about 5 mg/kg, about 7.5 kg/kg, or about 10 mg/kg is administered to the subject. In some embodiments, the antibody that specifically binds to the extracellular domain of human TIGIT is administered about once a week, about once every two weeks, about once every three weeks, or about once every four weeks, administered. In some embodiments, the subject has a histologically confirmed advanced recurrent or refractory solid tumor.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1002]
ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量をPD-1アンタゴニストまたはPD-L1アンタゴニストと組み合わせて対象に投与する工程を含む、該対象において腫瘍成長を阻害する方法であって、
該腫瘍が、高頻度マイクロサテライト不安定性結腸直腸がん、マイクロサテライト安定性結腸直腸がん、トリプルネガティブ乳がん、メルケル細胞がん、子宮内膜がん、または食道がんであり、かつ
ヒトTIGITに結合する該抗体が、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1003]
ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量を対象に投与する工程を含む、該対象において腫瘍成長を阻害する方法であって、
該腫瘍が、PD-1アンタゴニストまたはPD-L1アンタゴニストを用いる処置に対して抵抗性または不応性であり、かつ
ヒトTIGITに結合する該抗体が、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1004]
ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量をPD-1アンタゴニストまたはPD-L1アンタゴニストと組み合わせて対象に投与する工程を含む、該対象において腫瘍成長を阻害する方法であって、
該腫瘍が、単一の作用物質としてPD-1アンタゴニストまたはPD-L1アンタゴニストを用いる処置に対して抵抗性または不応性であり、かつ
ヒトTIGITに結合する該抗体が、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1005]
ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量を対象に投与する工程を含む、該対象において腫瘍成長を阻害する方法であって、
該対象が、単一の作用物質としてPD-1アンタゴニストまたはPD-L1アンタゴニストを用いて以前に処置されたことがあり、かつ
ヒトTIGITに結合する該抗体が、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1006]
ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量をPD-1アンタゴニストまたはPD-L1アンタゴニストと組み合わせて対象に投与する工程を含む、該対象において腫瘍成長を阻害する方法であって、
該対象が、単一の作用物質としてPD-1アンタゴニストまたはPD-L1アンタゴニストを用いて以前に処置されたことがあり、かつ
ヒトTIGITに結合する該抗体が、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1007]
ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量を対象に投与する工程を含む、該対象において腫瘍成長を阻害する方法であって、
該腫瘍がポリオウイルス受容体(PVR)および/またはポリオウイルス受容体関連2(PVRL2)を発現し、かつ
ヒトTIGITに結合する該抗体が、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1008]
ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量をPD-1アンタゴニストまたはPD-L1アンタゴニストと組み合わせて対象に投与する工程を含む、該対象において腫瘍成長を阻害する方法であって、
該腫瘍が、ポリオウイルス受容体(PVR)および/またはポリオウイルス受容体関連2(PVRL2)を発現し、かつ
ヒトTIGITに結合する該抗体が、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1009]
ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量を対象に投与する工程を含む、該対象において腫瘍成長を阻害する方法であって、
該腫瘍が腫瘍浸潤リンパ球(TIL)を含み、かつ
ヒトTIGITに結合する該抗体が、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1010]
ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量をPD-1アンタゴニストまたはPD-L1アンタゴニストと組み合わせて対象に投与する工程を含む、該対象において腫瘍成長を阻害する方法であって、
該腫瘍が腫瘍浸潤リンパ球を含み、かつ
ヒトTIGITに結合する該抗体が、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1011]
ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量を対象に投与する工程を含む、該対象において腫瘍成長を阻害する方法であって、
該腫瘍が調節性T細胞を含み、かつ
ヒトTIGITに結合する該抗体が、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1012]
ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量をPD-1アンタゴニストまたはPD-L1アンタゴニストと組み合わせて対象に投与する工程を含む、該対象において腫瘍成長を阻害する方法であって、
該腫瘍が調節性T細胞を含み、かつ
ヒトTIGITに結合する該抗体が、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1013]
PD-1アンタゴニストまたはPD-L1アンタゴニストを用いる処置の最中または後に腫瘍成長進行がある、本発明1003～1006のいずれかの方法。
[本発明1014]
PD-1アンタゴニストまたはPD-L1アンタゴニストを用いる処置の後に腫瘍成長再発がある、本発明1003～1006のいずれかの方法。
[本発明1015]
前記腫瘍再発が局所再発、局部再発、または遠隔再発である、本発明1014の方法。
[本発明1016]
前記腫瘍が、抗PD-1抗体を用いる処置に対して抵抗性または不応性である、本発明1003または本発明1004の方法。
[本発明1017]
前記腫瘍が、抗PD-L1抗体を用いる処置に対して抵抗性または不応性である、本発明1003または本発明1004の方法。
[本発明1018]
前記対象が、抗PD-1抗体を用いて以前に処置されたことがある、本発明1005または本発明1006の方法。
[本発明1019]
前記対象が、抗PD-L1抗体を用いて以前に処置されたことがある、本発明1005または本発明1006の方法。
[本発明1020]
併用療法が抗PD-1抗体を含む、本発明1002、1004、1006、1008、1010、または1012のいずれかの方法。
[本発明1021]
併用療法が抗PD-L1抗体を含む、本発明1002、1004、1006、1008、1010、または1012のいずれかの方法。
[本発明1022]
前記腫瘍が、肺腫瘍、肝臓腫瘍、乳房腫瘍、腎細胞がん/腎臓腫瘍、前立腺腫瘍、胃腸腫瘍/胃腫瘍、黒色腫、頸部腫瘍、膀胱腫瘍、グリア芽細胞腫、頭頸部腫瘍、膵臓腫瘍、卵巣腫瘍、結腸直腸腫瘍、子宮内膜腫瘍、および食道腫瘍からなる群より選択される、本発明1003～1021のいずれかの方法。
[本発明1023]
前記ヒトTIGITに結合する抗体が、SEQ ID NO:10を含む重鎖可変領域、およびSEQ ID NO:11を含む軽鎖可変領域を含む、本発明1001～1022のいずれかの方法。
[本発明1024]
前記ヒトTIGITに結合する抗体がモノクローナル抗体である、本発明1001～1023のいずれかの方法。
[本発明1025]
前記ヒトTIGITに結合する抗体が二重特異性抗体である、本発明1001～1024のいずれかの方法。
[本発明1026]
前記ヒトTIGITに結合する抗体が、抗原結合部位を含む抗体断片である、本発明1001～1025のいずれかの方法。
[本発明1027]
前記ヒトTIGITに結合する抗体がIgG1抗体である、本発明1001～1025のいずれかの方法。
[本発明1028]
前記ヒトTIGITに結合する抗体がIgG4抗体である、本発明1001～1025のいずれかの方法。
[本発明1029]
前記ヒトTIGITに結合する抗体が、SEQ ID NO:13の重鎖アミノ酸配列、およびSEQ ID NO:15の軽鎖アミノ酸配列を含む、本発明1001～1025のいずれかの方法。
[本発明1030]
前記ヒトTIGITに結合する抗体が、SEQ ID NO:17の重鎖アミノ酸配列、およびSEQ ID NO:15の軽鎖アミノ酸配列を含む、本発明1001～1025のいずれかの方法。
[本発明1031]
前記ヒトTIGITに結合する抗体が、指定番号PTA-122346としてATCCに寄託されたプラスミドによってコードされる重鎖可変領域、および指定番号PTA-122347としてATCCに寄託されたプラスミドによってコードされる軽鎖可変領域を含む、本発明1001～1026のいずれかの方法。
[本発明1032]
前記腫瘍内および/または腫瘍微小環境内のTエフェクター細胞を増大させる、本発明1001～1031のいずれかの方法。
[本発明1033]
前記腫瘍内および/または腫瘍微小環境内のTエフェクター細胞活性を増大させる、本発明1001～1031のいずれかの方法。
[本発明1034]
前記腫瘍内および/または腫瘍微小環境内の調節性T細胞を枯渇させる、本発明1001～1031のいずれかの方法。
[本発明1035]
前記腫瘍内および/または腫瘍微小環境内のTregの抑制活性を阻害しかつ/または減少させる、本発明1001～1031のいずれかの方法。
[本発明1036]
前記ヒトTIGITに結合する抗体が、週1回、2週間に1回、3週間に1回、または4週間に1回投与される、本発明1001～1035のいずれかの方法。
[本発明1037]
前記ヒトTIGITに結合する抗体が、約2週間に1回または3週間に1回投与される、本発明1036の方法。
[本発明1038]
前記ヒトTIGITに結合する抗体が、約0.1mg/kg～約25mg/kgの用量で投与される、本発明1001～1037のいずれかの方法。
[本発明1039]
前記ヒトTIGITに結合する抗体が、約0.3mg/kg、約1mg/kg、約3mg/kg、約10mg/kg、または約15mg/kgの用量で投与される、本発明1038の方法。
[本発明1040]
少なくとも1種類のさらなる治療用物質を投与する工程をさらに含む、本発明1001～1039のいずれかの方法。
[本発明1041]
前記さらなる治療用物質が化学療法剤である、本発明1040の方法。
[本発明1042]
前記さらなる治療用物質が抗体である、本発明1040の方法。
[本発明1043]
前記さらなる治療用物質が免疫療法剤である、本発明1040の方法。
[本発明1044]
前記免疫療法剤が、GM-CSF、M-CSF、G-CSF、IL-2、IL-3、IL-12、IL-15、B7-1(CD80)、B7-2(CD86)、4-1BBリガンド、GITRL、OX40L、CD40L、抗CD3抗体、抗CTLA-4抗体、抗GITR抗体、抗OX40抗体、抗4-1BB抗体、抗LAG-3抗体、および抗TIM-3抗体からなる群より選択される、本発明1043の方法。
[本発明1045]
前記さらなる治療用物質が、Notch経路、Wnt経路、またはRSPO/LGR経路の阻害物質である、本発明1040の方法。
[本発明1046]
前記対象の腫瘍またはがんが少なくとも部分的に除去されている、本発明1001～1045のいずれかの方法。
[本発明1047]
ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量を対象に投与する工程を含む、該対象において腫瘍成長を阻害する方法であって、
該腫瘍が、頭頸部がん、食道がん/胃食道がん、胃がん、結腸直腸がん、肛門がん、肝細胞がん/肝臓がん、子宮頸がん、肺がん、黒色腫、メルケル細胞がん、腎細胞がん/腎臓がん、膀胱がん、卵巣がん、膵臓がん、子宮内膜がん、およびトリプルネガティブ乳がん、既知高頻度MSI固形腫瘍(MSI CRCを含む)またはMSS結腸直腸がんであり、
該腫瘍が、抗PD-1抗体または抗PD-L1抗体を用いる処置に対して抵抗性または不応性であり、かつ
ヒトTIGITに結合する該抗体が、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、該方法。
[本発明1048]
前記対象が、抗PD-1抗体または抗PD-L1抗体を用いて以前に処置されたことがある、本発明1047の方法。
[本発明1049]
前記ヒトTIGITの細胞外ドメインに特異的に結合する抗体が、約0.1mg/kg、約0.3mg/kg、約1mg/kg、約1.5mg/kg、約2mg/kg、約2.5mg/kg、約3mg/kg、約5mg/kg、約7.5kg/kg、約10mg/kg、または約15mg/kgの用量で前記対象に投与される、本発明1001～1048のいずれかの方法。
[本発明1050]
前記ヒトTIGITの細胞外ドメインに特異的に結合する抗体が、週1回、2週間に1回、3週間に1回、または4週間に1回投与される、本発明1001～1049のいずれかの方法。
[本発明1051]
前記対象が、組織学的に確認された進行型の再発性または不応性固形腫瘍を有する、本発明1001～1050のいずれかの方法。
Where aspects or embodiments of the invention have been described in Markush groups or other groups of alternatives, the invention includes not only the recited group as a whole, but also each member of the group individually, including the main group Also includes all possible subgroups of , including main groups where one or more of the group members are absent. The present invention also contemplates the explicit exclusion of one or more of any group members in the present invention.
[Invention 1001]
1. A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, comprising:
the tumor is microsatellite instability-high colorectal cancer, microsatellite stable colorectal cancer, triple-negative breast cancer, Merkel cell carcinoma, endometrial cancer, or have esophageal cancer, and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1002]
A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or PD-L1 antagonist. hand,
the tumor is high-frequency microsatellite-unstable colorectal cancer, microsatellite-stable colorectal cancer, triple-negative breast cancer, Merkel cell carcinoma, endometrial cancer, or esophageal cancer, and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1003]
1. A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, comprising:
the tumor is resistant or refractory to treatment with a PD-1 antagonist or PD-L1 antagonist, and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1004]
A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or PD-L1 antagonist. hand,
the tumor is resistant or refractory to treatment with a PD-1 antagonist or PD-L1 antagonist as a single agent, and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1005]
1. A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, comprising:
The subject has been previously treated with a PD-1 antagonist or a PD-L1 antagonist as a single agent, and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1006]
A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or PD-L1 antagonist. hand,
The subject has been previously treated with a PD-1 antagonist or a PD-L1 antagonist as a single agent, and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1007]
1. A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, comprising:
the tumor expresses poliovirus receptor (PVR) and/or poliovirus receptor-related 2 (PVRL2), and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1008]
A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or PD-L1 antagonist. hand,
the tumor expresses poliovirus receptor (PVR) and/or poliovirus receptor-related 2 (PVRL2), and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1009]
1. A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, comprising:
the tumor comprises tumor infiltrating lymphocytes (TILs), and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1010]
A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or PD-L1 antagonist. hand,
the tumor comprises tumor-infiltrating lymphocytes, and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1011]
1. A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, comprising:
the tumor comprises regulatory T cells, and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1012]
A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT in combination with a PD-1 antagonist or PD-L1 antagonist. hand,
the tumor comprises regulatory T cells, and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1013]
1006. The method of any of inventions 1003-1006, wherein there is tumor growth progression during or after treatment with a PD-1 antagonist or PD-L1 antagonist.
[Invention 1014]
1006. The method of any of inventions 1003-1006, wherein there is tumor growth recurrence following treatment with a PD-1 antagonist or PD-L1 antagonist.
[Invention 1015]
1015. The method of invention 1014, wherein said tumor recurrence is local recurrence, regional recurrence, or distant recurrence.
[Invention 1016]
The method of invention 1003 or invention 1004, wherein said tumor is resistant or refractory to treatment with an anti-PD-1 antibody.
[Invention 1017]
The method of invention 1003 or invention 1004, wherein said tumor is resistant or refractory to treatment with an anti-PD-L1 antibody.
[Invention 1018]
The method of invention 1005 or invention 1006, wherein said subject has been previously treated with an anti-PD-1 antibody.
[Invention 1019]
The method of invention 1005 or invention 1006, wherein said subject has been previously treated with an anti-PD-L1 antibody.
[Invention 1020]
The method of any of invention 1002, 1004, 1006, 1008, 1010, or 1012, wherein the combination therapy comprises an anti-PD-1 antibody.
[Invention 1021]
The method of any of invention 1002, 1004, 1006, 1008, 1010, or 1012, wherein the combination therapy comprises an anti-PD-L1 antibody.
[Invention 1022]
said tumor is lung tumor, liver tumor, breast tumor, renal cell carcinoma/kidney tumor, prostate tumor, gastrointestinal/stomach tumor, melanoma, neck tumor, bladder tumor, glioblastoma, head and neck tumor, pancreas The method of any of the inventions 1003-1021 selected from the group consisting of tumor, ovarian tumor, colorectal tumor, endometrial tumor, and esophageal tumor.
[Invention 1023]
The method of any of inventions 1001-1022, wherein said antibody that binds to human TIGIT comprises a heavy chain variable region comprising SEQ ID NO:10 and a light chain variable region comprising SEQ ID NO:11.
[Invention 1024]
The method of any of inventions 1001-1023, wherein said antibody that binds to human TIGIT is a monoclonal antibody.
[Invention 1025]
The method of any of inventions 1001-1024, wherein said antibody that binds to human TIGIT is a bispecific antibody.
[Invention 1026]
The method of any of inventions 1001-1025, wherein said antibody that binds to human TIGIT is an antibody fragment comprising an antigen binding site.
[Invention 1027]
The method of any of inventions 1001-1025, wherein said antibody that binds to human TIGIT is an IgG1 antibody.
[Invention 1028]
The method of any one of inventions 1001-1025, wherein said antibody that binds to human TIGIT is an IgG4 antibody.
[Invention 1029]
The method of any of inventions 1001-1025, wherein said antibody that binds to human TIGIT comprises the heavy chain amino acid sequence of SEQ ID NO:13 and the light chain amino acid sequence of SEQ ID NO:15.
[Invention 1030]
The method of any of inventions 1001-1025, wherein said antibody that binds to human TIGIT comprises the heavy chain amino acid sequence of SEQ ID NO:17 and the light chain amino acid sequence of SEQ ID NO:15.
[Invention 1031]
wherein said antibody that binds to human TIGIT is a heavy chain variable region encoded by a plasmid deposited with ATCC under designation number PTA-122346 and a light chain variable region encoded by a plasmid deposited with ATCC under designation number PTA-122347; 1026. The method of any of the inventions 1001-1026 comprising a region.
[Invention 1032]
The method of any of the inventions 1001-1031, wherein T effector cells within said tumor and/or within the tumor microenvironment are expanded.
[Invention 1033]
The method of any of invention 1001-1031, wherein T effector cell activity within said tumor and/or within the tumor microenvironment is increased.
[Invention 1034]
The method of any of the inventions 1001-1031, wherein regulatory T cells within said tumor and/or within the tumor microenvironment are depleted.
[Invention 1035]
The method of any of the inventions 1001-1031, wherein the suppressive activity of Tregs within said tumor and/or tumor microenvironment is inhibited and/or reduced.
[Invention 1036]
The method of any of inventions 1001-1035, wherein said antibody that binds to human TIGIT is administered once a week, once every two weeks, once every three weeks, or once every four weeks.
[Invention 1037]
1036. The method of invention 1036, wherein said antibody that binds to human TIGIT is administered about once every two weeks or once every three weeks.
[Invention 1038]
The method of any of inventions 1001-1037, wherein said antibody that binds to human TIGIT is administered at a dose of about 0.1 mg/kg to about 25 mg/kg.
[Invention 1039]
1038. The method of invention 1038, wherein said antibody that binds to human TIGIT is administered at a dose of about 0.3 mg/kg, about 1 mg/kg, about 3 mg/kg, about 10 mg/kg, or about 15 mg/kg.
[Invention 1040]
The method of any of inventions 1001-1039, further comprising administering at least one additional therapeutic agent.
[Invention 1041]
The method of invention 1040, wherein said additional therapeutic agent is a chemotherapeutic agent.
[Invention 1042]
The method of invention 1040, wherein said additional therapeutic agent is an antibody.
[Invention 1043]
The method of invention 1040, wherein said additional therapeutic agent is an immunotherapeutic agent.
[Invention 1044]
the immunotherapeutic agent is GM-CSF, M-CSF, G-CSF, IL-2, IL-3, IL-12, IL-15, B7-1 (CD80), B7-2 (CD86), 4- selected from the group consisting of 1BB ligand, GITRL, OX40L, CD40L, anti-CD3 antibody, anti-CTLA-4 antibody, anti-GITR antibody, anti-OX40 antibody, anti-4-1BB antibody, anti-LAG-3 antibody, and anti-TIM-3 antibody method of the present invention 1043.
[Invention 1045]
1040. The method of invention 1040, wherein said additional therapeutic agent is an inhibitor of the Notch pathway, Wnt pathway, or RSPO/LGR pathway.
[Invention 1046]
The method of any of inventions 1001-1045, wherein the subject's tumor or cancer is at least partially ablated.
[Invention 1047]
1. A method of inhibiting tumor growth in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT, comprising:
The tumor is head and neck cancer, esophageal cancer/gastroesophageal cancer, gastric cancer, colorectal cancer, anal cancer, hepatocellular carcinoma/liver cancer, cervical cancer, lung cancer, melanoma, Merkel cell Cancer, renal cell/kidney cancer, bladder cancer, ovarian cancer, pancreatic cancer, endometrial cancer, and triple-negative breast cancer, known MSI solid tumors (including MSI CRC) or MSS colon have rectal cancer,
the tumor is resistant or refractory to treatment with an anti-PD-1 antibody or an anti-PD-L1 antibody, and
the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).
[Invention 1048]
1047. The method of invention 1047, wherein said subject has been previously treated with an anti-PD-1 antibody or an anti-PD-L1 antibody.
[Invention 1049]
about 0.1 mg/kg, about 0.3 mg/kg, about 1 mg/kg, about 1.5 mg/kg, about 2 mg/kg, about 2.5 mg/kg, The method of any of inventions 1001-1048, wherein the subject is administered a dose of about 3 mg/kg, about 5 mg/kg, about 7.5 kg/kg, about 10 mg/kg, or about 15 mg/kg.
[Invention 1050]
Any of invention 1001-1049, wherein said antibody that specifically binds to the extracellular domain of human TIGIT is administered once a week, once every two weeks, once every three weeks, or once every four weeks the method of.
[Invention 1051]
The method of any of inventions 1001-1050, wherein said subject has a histologically confirmed advanced recurrent or refractory solid tumor.

Inhibition of tumor growth by the anti-TIGIT antibody OMP-313M32 in a humanized mouse model. Humanized mice were injected subcutaneously with patient-derived melanoma tumor cells (OMP-M9, 75,000 cells/mouse). Tumors were grown for 19 days until the average tumor volume was approximately 50 mm ³ . Tumor-bearing mice were randomized into groups (n=8 mice/group). Tumor-bearing mice were treated with control antibody or anti-TIGIT antibody OMP-313M32. Mice were dosed with 1 mg/kg or 5 mg/kg every 5 days. Tumor growth was monitored and tumor volumes were measured using electronic calipers at the indicated time points. Schematic showing dose escalation and escalation for early phase clinical trials with anti-TIGIT antibody 313M32.

DETAILED DESCRIPTION OF THE INVENTION The present invention provides methods of using novel agents including, but not limited to, polypeptides, antibodies, heterodimeric and homodimeric molecules that specifically bind to human TIGIT. offer. Methods of using the novel agents to inhibit tumor growth and/or treat cancer are provided. Methods of using novel agents, e.g., methods of activating an immune response, methods of stimulating an immune response, methods of enhancing an immune response, methods of increasing an immune response, natural killer (NK) cells and/or T Methods of activating cells, methods of increasing NK cell and/or T cell activity, methods of promoting NK cell and/or T cell activity, decreasing suppressor T cells (i.e. regulatory T cells) and Further provided are methods of inhibiting and/or reducing and/or inhibiting myeloid-derived suppressor cells.

I. Definitions To facilitate understanding of the present invention, a number of terms and phrases are defined below.

As used herein, the terms "agonist" and "agonistic" directly or indirectly substantially induce, activate or promote the biological activity of a target and/or pathway refers to or describes an agent that can be increased or enhanced. The term "agonist" is used herein to include any agent that partially or fully induces, activates, promotes, increases or enhances the activity of a protein. be done.

As used herein, the terms "antagonist" and "antagonistic" are intended to directly or indirectly partially or completely block, inhibit, or reduce the biological activity of a target and/or pathway. refers to or describes an agent that can be neutralized or neutralized. The term "antagonist" is used herein to include any molecule that partially or fully blocks, inhibits, reduces or neutralizes the activity of a protein.

The terms "modulate" and "modulate" as used herein refer to alteration or change in biological activity. Modulation includes, but is not limited to stimulating or inhibiting activity. A modulation can be an increase or decrease in activity, an alteration in binding properties, or a biological, functional, or immunological property related to the activity of a protein, pathway, system, or other biological target of interest. Any other change in the physical characteristics is also possible.

The term "antibody" as used herein refers to an immunoglobulin molecule that recognizes and specifically binds to a target through at least one antigen binding site. Said target can be a protein, polypeptide, peptide, carbohydrate, polynucleotide, lipid, or any combination of the above. As used herein, the term includes intact polyclonal antibodies, intact monoclonal antibodies, antibody fragments (e.g., Fab, Fab', F(ab')2, and Fv fragments), single-chain Fv (scFv) Antibodies, multispecific antibodies, bispecific antibodies, monospecific antibodies, monovalent antibodies, chimeric antibodies, humanized antibodies, human antibodies, fusion proteins comprising antigen-binding portions of antibodies, as well as biological antibodies for which antibodies are desirable Any other modified immunoglobulin molecule containing an antigen binding site is included, so long as it exhibits activity. Antibodies are divided into five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, or their subclasses ( isotype) (eg, IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2). Different classes of immunoglobulins have different well-known subunit and three-dimensional structures. Antibodies can be naked or conjugated to other molecules including, but not limited to, toxins and radioisotopes.

The term "antibody fragment" refers to a portion of an intact antibody, and generally to the antigen-determining variable region of the intact antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2, and Fv fragments, linear antibodies, single-chain antibodies, and multispecific antibodies formed from antibody fragments. not. As used herein, an "antibody fragment" comprises an antigen binding site or epitope binding site.

The term "variable region" of an antibody refers to the variable region of the antibody light chain or the variable region of the antibody heavy chain, alone or in combination. Generally, a heavy or light chain variable region consists of four framework regions connected by three complementarity determining regions (CDRs), also known as "hypervariable regions". The CDRs within each chain are held together by framework regions and, together with the CDRs from other chains, contribute to the formation of the antibody's antigen-binding site. There are at least two techniques for determining CDRs: (1) approaches based on interspecies sequence variability (i.e., Kabat et al., Sequences of Proteins of Immunological Interest, 5th Edition., 1991, National Institutes of Health, Bethesda Md. )); and (2) an approach based on crystallographic studies of antigen-antibody complexes (Al-Lazikani et al (1997) J. Mol. Biol. 273:927-948)). Additionally, a combination of these two approaches is sometimes used in the art to determine CDRs.

As used herein, the term "monoclonal antibody" refers to a homogeneous antibody population responsible for the highly specific recognition and binding of a single antigenic determinant, or epitope. This is in contrast to polyclonal antibodies, which typically include a mixture of different antibodies that recognize different antigenic determinants. The term "monoclonal antibody" includes intact and full-length monoclonal antibodies, as well as antibody fragments (e.g., Fab, Fab', F(ab')2, Fv), single chain (scFv) antibodies, antibody fragments and any other modified immunoglobulin molecule that contains an antigen binding site. Additionally, "monoclonal antibody" refers to such antibodies produced by any number of techniques including, but not limited to, hybridoma production, phage selection, recombinant expression, and transgenic animals.

As used herein, the term "humanized antibody" refers to antibodies that are specific immunoglobulin chains, chimeric immunoglobulins, or fragments thereof that contain minimal non-human sequences. Typically, humanized antibodies are derived from CDRs of a non-human species (e.g., mouse, rat, rabbit, or hamster) in which the amino acid residues of the CDRs have the desired specificity, affinity, and/or binding ability. A human immunoglobulin that has been replaced by amino acid residues. In some instances, framework variable region amino acid residues of the human immunoglobulin may be replaced by corresponding amino acid residues in an antibody from a non-human species. Furthermore, the humanized antibody is modified by substitution of additional amino acid residues in the framework variable region and/or within the replaced non-human amino acid residues to further improve antibody specificity, affinity, and/or binding. Capabilities can be refined and optimized. A humanized antibody may comprise a variable domain containing all or substantially all of the CDRs corresponding to a non-human immunoglobulin, whereas all or substantially all of the framework variable regions are Framework regions of human immunoglobulin sequences. In some embodiments, the variable domain comprises the framework regions of human immunoglobulin sequences. In some embodiments, the variable domain comprises framework regions of human immunoglobulin consensus sequences. The humanized antibody also will comprise at least a portion of an immunoglobulin constant region or domain (Fc), typically that of a human immunoglobulin constant region or domain (Fc).

As used herein, the term "human antibody" refers to an antibody produced by a human or an amino acid sequence corresponding to an antibody produced by a human produced using any technique known in the art. refers to an antibody that has

As used herein, the term "chimeric antibody" refers to an antibody in which the amino acid sequences of the immunoglobulin molecule are derived from two or more species. Typically, the light and heavy chain variable regions are antibody variable regions from one mammalian species (e.g., mouse, rat, rabbit, etc.) having the desired specificity, affinity, and/or binding ability. , while the constant regions are homologous to sequences in antibodies from different species.

The terms "epitope" or "antigenic determinant" are used interchangeably herein and refer to that portion of an antigen or target capable of being recognized and specifically bound by a particular antibody. When the antigen or target is a polypeptide, epitopes can be formed both from contiguous amino acids and noncontiguous amino acids juxtaposed by tertiary folding of the protein. Epitopes formed from contiguous amino acids (also called linear epitopes) are typically retained when the protein is denatured, whereas epitopes formed by tertiary folding (also called conformational epitopes) is typically lost when proteins are denatured. An epitope typically includes at least 3 amino acids, more usually at least 5, 6, 7, or 8-10 amino acids in a unique spatial arrangement.

The term "selectively binds" or "specifically binds" means that an agent binds more frequently, rapidly, over a longer period of time, with a higher affinity than another substance, including related and unrelated proteins. with or in combination with the above means reacting with an epitope, protein, or target molecule. In certain embodiments, "specifically binds" means that the agent binds to the protein or target, eg, with a K _D of about 0.1 mM or less, more usually less than about 1 μM. In certain embodiments, “specifically binds” means that the agent binds to the target with a K _D of at least about 0.1 μM or less, at least about 0.01 μM or less, or at least about 1 nM or less. means to Because there is sequence identity between homologous proteins in different species, specific binding may involve agents that recognize proteins or targets in multiple species (eg, mouse TIGIT and human TIGIT). Similarly, specific binding may involve agents that recognize more than one type of protein or target, since there is homology within certain regions of the polypeptide sequences of different proteins. It is understood that in certain embodiments, an agent that specifically binds to a first target may or may not specifically bind to a second target. Thus, "specific binding" does not necessarily require exclusive binding, ie, binding to one type of target (although it may include binding to one type of target). Thus, an agent may, in certain embodiments, specifically bind to more than one type of target. In certain embodiments, multiple types of targets may bind to the same antigen binding site of the agent. For example, an antibody may, in certain cases, contain two identical antigen binding sites, each of which specifically binds to the same epitope on two or more proteins. In certain other embodiments, antibodies may be bispecific, comprising at least two antigen binding sites of differing specificity. Generally, but not necessarily, references to binding imply specific binding.

As used herein, "selecting" and "selected" with respect to a patient are based on the particular patient having predetermined criteria (referred to as having predetermined criteria). reason), for example, to mean that this patient is specifically selected from a large group of patients based on having tumors with high levels of PVR and/or PVRL2 expression. Similarly, "selectively treating a patient with a tumor" refers to a particular patient based on (because of having predetermined criteria) having predetermined criteria, e.g., this It refers to providing treatment to cancer patients specifically selected from a large group of patients based on the patient having a tumor with high PVR and/or PVRL2 expression levels. Similarly, "selectively administering" is based on (because of having predetermined criteria) a particular patient has predetermined criteria, e.g., if the patient has PVR and / Or Refers to administering drugs to cancer patients specifically selected from a large group of patients based on having tumors with high levels of PVRL2 expression. Selecting, selectively treating, and selectively administering refer to the treatment of a patient rather than subjecting the patient to standard treatment regimens based solely on the fact that the patient has a cancer such as CRC or NSCLC. It is meant to provide patients with personalized cancer therapy based on cancer biology.

The terms "polypeptide" and "peptide" and "protein" are used interchangeably herein to refer to polymers of amino acids of any length. The polymer may be linear or branched, may contain modified amino acids, and may be interrupted by non-amino acids. These terms are also modified naturally or by intervention, for example by disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation, or any other manipulation or modification such as conjugation with a labeling component. Also included are amino acid polymers that have been modified. For example, polypeptides containing one or more amino acid analogs (eg, including unnatural amino acids) as well as other modifications known in the art are also included in this definition. Since the polypeptides of the invention may be based in certain embodiments on antibodies or other immunoglobulin superfamily members, it is understood that a "polypeptide" can occur as a single chain or two or more associated chains. understood.

The terms "polynucleotide" and "nucleic acid" and "nucleic acid molecule" are used interchangeably herein to refer to polymers of nucleotides of any length, and include DNA and RNA. Nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and/or analogues thereof, or any substrate that can be incorporated into a polymer by a DNA or RNA polymerase.

The terms "identical" or percent "identity" in the context of two or more nucleic acids or polypeptides are intended to give maximum correspondence without considering any conservative amino acid substitutions as part of the sequence identity. refers to two or more sequences or subsequences that are the same or have a certain percentage of the same nucleotides or amino acid residues when compared and aligned (with gaps introduced if necessary) to . Percent identity may be determined using sequence comparison software or algorithms, or by visual inspection. Various algorithms and software that may be used to obtain alignments of amino acid or nucleotide sequences are well known in the art. These algorithms and software include, but are not limited to BLAST, ALIGN, Megalign, BestFit, GCG Wisconsin Package, and variations thereof. In some embodiments, two nucleic acids or polypeptides of the invention are substantially identical, and substantially identical are compared and aligned for maximum correspondence between the two nucleic acids or polypeptides. at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, in some embodiments at least 95%, 96%, when determined using a sequence comparison algorithm or by visual inspection. %, 97%, 98%, 99% nucleotide or amino acid residue identity. In some embodiments, identity is at least about 10, at least about 20, at least about 40-60 nucleotides or amino acid residues, at least about 60-80 nucleotides or amino acid residues in length, or Reside over the region of the array, any integer value in between. In some embodiments, the identity exists over a region longer than 60-80 nucleotides or amino acid residues, such as at least about 80-100 nucleotides or amino acid residues; are substantially identical over the entire length of the coding regions of the sequences being compared, eg, the nucleotide sequences.

A "conservative amino acid substitution" is one in which one amino acid residue is replaced with another amino acid residue having a similar side chain. Generally, basic side chains (e.g. lysine, arginine, histidine), acidic side chains (e.g. aspartic acid, glutamic acid), uncharged polar side chains (e.g. glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), non-polar side chains (e.g. alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), β-branched side chains (e.g. threonine, valine, isoleucine), and aromatic side chains (e.g. Families of amino acid residues with similar side chains have been defined in the art, including tyrosine, phenylalanine, tryptophan, histidine). For example, the substitution of phenylalanine for tyrosine is contemplated as a conservative substitution. Generally, conservative substitutions in the sequences of the polypeptides and/or antibodies of the invention do not prevent binding of the polypeptide or antibody containing the amino acid sequence to the target binding site. Methods for identifying conservative substitutions of nucleotides and amino acids that do not eliminate binding are well known in the art.

As used herein, the term "vector" refers to a construct, usually expressible, capable of delivering one or more genes or sequences of interest into a host cell. Examples of vectors include viral vectors, naked DNA or RNA expression vectors, plasmid, cosmid, or phage vectors, DNA or RNA expression vectors conjugated with cationic condensing agents, and DNA encapsulated in liposomes. Including, but not limited to, expression vectors or RNA expression vectors.

An "isolated" polypeptide, soluble protein, antibody, polynucleotide, vector, cell, or composition is a polypeptide, soluble protein, antibody, polynucleotide, vector, cell, or composition in a form not found in nature. or a composition. An isolated polypeptide, soluble protein, antibody, polynucleotide, vector, cell, or composition is a polypeptide, soluble protein, antibody, polynucleotide that has been purified to the extent that it is no longer in the form in which it is found in nature. , vectors, cells or compositions. In some embodiments, an isolated polypeptide, soluble protein, antibody, polynucleotide, vector, cell, or composition is substantially pure.

As used herein, the term "substantially pure" means at least 50% pure (i.e. free of contaminants), at least 90% pure, at least 95% pure, at least 98% pure, Or refers to materials that are at least 99% pure.

As used herein, the term "immune response" includes responses from both the innate and adaptive immune system. An "immune response" includes both cellular and/or humoral immune responses. An "immune response" includes, but is not limited to, both T and B cell responses, as well as responses from other immune system cells such as natural killer (NK) cells, monocytes, macrophages, and the like.

The terms "cancer" and "cancerous" as used herein refer to or describe the physiological condition in mammals having cell populations characterized by unregulated cell growth. Examples of cancer include, but are not limited to, carcinoma, blastoma, sarcoma, and hematologic cancers such as lymphoma and leukemia.

As used herein, the terms "tumor" and "neoplasm" are either benign (non-cancerous) or malignant (cancerous), including precancerous lesions, resulting from excessive cell growth or proliferation. refers to any tissue mass in

As used herein, the term "metastasis" refers to the process by which cancer spreads or migrates from its site of origin to other body areas, resulting in the development of similar cancer lesions at new locations. point to In general, a "metastatic" or "metastatic" cell is one that has lost adherent contacts with neighboring cells and has traveled through the body from the primary site of disease via the bloodstream or lymph to secondary sites. cells that migrate to

The terms "cancer cell" and "tumor cell" refer to cancers or tumors or progenitor cells, including both non-tumorigenic cells and tumorigenic stem cells (cancer stem cells), which make up the majority of the cancer cell population. Refers to the entire population of cells derived from cancerous lesions. As used herein, the term "cancer cell" or "tumor cell" refers only to cells that are incapable of replication and differentiation, to distinguish such tumor cells from cancer stem cells. modified by the term "non-tumorigenic".

The term "subject" is any animal (e.g., mammal) that is the recipient of a particular treatment, including but not limited to humans, non-human primates, dogs, cats, rabbits, rodents, and the like. point to Typically, the terms "subject" and "patient" are used interchangeably herein with respect to human subjects.

The term "pharmaceutically acceptable" means a substance that has been approved or sanctioned by a federal or state regulatory agency, or the United States Pharmacopoeia, or a generally accepted substance for use in animals, including humans. Refers to substances listed in other pharmacopoeias

The term "pharmaceutically acceptable excipient, carrier, or adjuvant" or "acceptable pharmaceutical carrier" can be administered to a subject with at least one agent of the disclosure, and the pharmacological Refers to an excipient, carrier, or adjuvant that is non-toxic when administered in doses sufficient to produce a therapeutic effect without destroying its therapeutic activity. Generally, those skilled in the art and the US Food and Drug Administration consider pharmaceutically acceptable excipients, carriers, or adjuvants to be inactive ingredients of any formulation.

The terms "effective amount" or "therapeutically effective amount" or "therapeutic effect" refer to an agent, antibody, polypeptide, polynucleotide, Refers to the amount of small organic molecules, or other drugs. In the case of cancer or tumors, a therapeutically effective amount of an agent (e.g., polypeptide or antibody) has a therapeutic effect and thus may enhance or boost an immune response or induce an anti-tumor response. may enhance or boost; may increase the cytolytic activity of immune cells; may increase killing of tumor cells; may increase killing of tumor cells by immune cells; may reduce the number of tumor cells, may reduce tumorigenicity, tumorigenic frequency, or tumorigenic potential; may reduce the number or frequency of cancer stem cells; may reduce tumor size; reduce cancer cell populations, inhibit or stop cancer cell infiltration into peripheral organs, including, for example, the spread of cancer to soft tissue and bone may inhibit and arrest tumor or cancer cell metastasis, inhibit and arrest tumor or cancer cell growth, or alleviate to some extent one or more of the symptoms associated with cancer. may reduce morbidity and mortality, may improve quality of life, or may be a combination of such effects.

The terms "treat" or "treatment" or "to treat" or "alleviate" or "to alleviate" are defined as (1) curing the symptoms of a diagnosed pathological condition or disorder, A therapeutic treatment that delays symptoms of an diagnosed pathological condition or disorder, relieves symptoms of a diagnosed pathological condition or disorder, and/or arrests progression of a diagnosed pathological condition or disorder, and ( 2) Refers to both prophylactic or preventative treatment that prevents or delays the onset of a targeted pathological condition or disorder. Thus, those in need of treatment include those who already have the disorder; those who are susceptible to the disorder; and those who seek to prevent the disorder. In the case of cancer or tumor, a subject has been successfully "treated" according to the methods of the invention if the patient exhibits one or more of the following: increased immune response, increased anti-tumor response, increased immune Increased cytolytic activity of cells, increased tumor cell killing, increased tumor cell killing by immune cells, reduced number or complete absence of cancer cells, reduced tumor size, soft tissue and bone inhibition or lack of cancer cell invasion to peripheral organs, including cancer cell expansion; inhibition or lack of tumor or cancer cell metastasis; inhibition or lack of cancer growth; Relief of multiple symptoms, reduced morbidity and mortality, improved quality of life, reduced tumorigenicity, reduced number or frequency of cancer stem cells, or any combination of effects.

As used in this disclosure and claims, the singular forms "a," "an," and "the" include plural forms unless the context clearly dictates otherwise. .

Wherever an embodiment is described herein with the term "comprising," other similar embodiments described with the term "consisting of" and/or "consisting essentially of" are also provided. is understood. It is also understood that whenever an aspect is described herein with the words “consisting essentially of”, other similar aspects described with “consisting of” are also provided.

As used herein, reference to "about" a value or parameter or "about" a value or parameter includes (and describes) aspects that are directed to that value or parameter. For example, description referring to "about X" includes description of "X."

The term "and/or" as used herein in phrases such as "A and/or B" is intended to include A and B, A or B, A (only), and B (only). be. Similarly, the term "and/or" used in phrases such as "A, B, and/or C" is intended to encompass each of the following aspects: A, B, and C; A and C; A and B; B and C; A (only); B (only); and C (only).

II. Methods of Use and Pharmaceutical Compositions The TIGIT binding agents of the present invention are useful in a variety of applications including, but not limited to, therapeutic treatment methods such as cancer treatment methods. In some embodiments, the therapeutic treatment method comprises immunotherapy against cancer. In certain embodiments, the TIGIT binding agent is useful for activating, promoting, increasing and/or enhancing an immune response, inhibiting tumor growth, reducing tumor volume, inhibiting tumor cell apoptosis. and/or to reduce the tumorigenicity of tumors.

The present invention provides methods for inhibiting tumor growth using TIGIT binding agents. In certain embodiments, the method of inhibiting tumor growth comprises administering a therapeutically effective amount of a TIGIT binding agent to the subject. In some embodiments, the TIGIT binding agent is an antibody that specifically binds to the extracellular domain of human TIGIT. In certain embodiments, the subject is human. In certain embodiments, the subject has a tumor or the subject has had a tumor at least partially removed.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、方法を提供する。一部の態様において、腫瘍は食道がんである。一部の態様において、腫瘍は、切除不能な、および/または転移性の疾患に対する少なくとも1つの先の全身療法または治療方針の際に進行していた食道がんである。一部の態様において、腫瘍はトリプルネガティブ乳がんである。一部の態様において、腫瘍は、不治の、進行型のエストロゲン受容体陰性、プロゲステロン受容体陰性、およびヒト上皮増殖因子受容体2陰性乳腺がんとして組織学的に確認されているトリプルネガティブ乳がんである。 In some embodiments, the invention provides a method of inhibiting tumor growth (eg, treating cancer) in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent, wherein the tumor is , microsatellite unstable colorectal cancer (MSI CRC), microsatellite stable colorectal cancer (MSS CRC), triple negative breast cancer (TNBC), Merkel cell carcinoma, endometrial cancer, or esophageal cancer I will provide a method. In some embodiments, a method of inhibiting tumor growth (e.g., treating cancer) in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT. and the tumor is microsatellite unstable colorectal cancer (MSI CRC), microsatellite stable colorectal cancer (MSS CRC), triple negative breast cancer (TNBC), Merkel cell carcinoma, endometrial cancer or esophageal cancer and the antibody that binds to human TIGIT has a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the tumor is esophageal cancer. In some embodiments, the tumor is esophageal cancer that has progressed during at least one prior systemic therapy or course of treatment for unresectable and/or metastatic disease. In some embodiments, the tumor is triple negative breast cancer. In some embodiments, the tumor is triple-negative breast cancer histologically confirmed as incurable, advanced estrogen receptor-negative, progesterone-receptor-negative, and human epidermal growth factor receptor 2-negative breast adenocarcinoma. be.

In some embodiments, histological confirmation is performed by one skilled in the art, eg, a trained clinician, using any method known in the art, eg, immunohistochemistry.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、方法を提供する。一部の態様において、高頻度MSI固形腫瘍には、MSI CRC、MSI胃がん、MSI子宮内膜がん、MSI卵巣がん、MSI肝胆道がん、MSI尿路がん、MSI脳がん、またはMSI皮膚がんが含まれ得る。 In some embodiments, the invention provides a method of inhibiting tumor growth (eg, treating cancer) in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent, wherein the tumor is A method is provided that is a high frequency MSI solid tumor. In some embodiments, a method of inhibiting tumor growth (e.g., treating cancer) in a subject comprising administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT. wherein the tumor is a high frequency MSI solid tumor and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the common MSI solid tumors include MSI CRC, MSI gastric cancer, MSI endometrial cancer, MSI ovarian cancer, MSI hepatobiliary cancer, MSI urinary tract cancer, MSI brain cancer, or MSI skin cancer may be included.

In general, tumors can be classified according to microsatellite stability (MSI). For example, solid tumors can be classified as MSI-high, MSI-low or MSI-stable tumors. MSI-high tumors can include, but are not limited to, colon cancer, stomach cancer, endometrial cancer, ovarian cancer, hepatobiliary cancer, urinary tract cancer, brain cancer, and skin cancer. not. MSI can be confirmed using any method known in the art, such as polymerase chain reaction (PCR) or immunohistochemistry (IHC). In some embodiments, MSI is determined in samples obtained from cancer patients. In some embodiments, the sample is a biopsy (eg, needle biopsy). In some embodiments, MSI includes, but is not limited to, reverse transcription PCR (RT-PCR), quantitative RT-PCR (qPCR), TaqMan™, or TaqMan™ low density array (TLDA). not determined in a polymerase chain reaction (PCR)-based method. In some embodiments, MSI is determined using a microarray. In some embodiments, MSI is determined by DNA sequencing, such as next generation sequencing. In some embodiments, MSI is determined in an immunohistochemistry (IHC) assay. In some embodiments, tumor MSI is compared to a previously determined MSI. In some embodiments, a tumor is classified as an MSI-high tumor if the MSI of the tumor is greater than a pre-determined MSI. In some embodiments, the predetermined MSI is the MSI of a reference tumor sample, a reference normal tissue sample, a series of reference tumor samples, or a series of reference normal tissue samples.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、対象において腫瘍成長を阻害する(例えば、がんを処置する)方法は、ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量をPD-L1アンタゴニストと組み合わせて対象に投与する工程を含み、腫瘍は、高頻度マイクロサテライト不安定性結腸直腸がん、マイクロサテライト安定性結腸直腸がん、トリプルネガティブ乳がん、メルケル細胞がん、子宮内膜がん、または食道がんであり、かつヒトTIGITに結合する抗体は、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、腫瘍は食道がんである。一部の態様において、腫瘍は、切除不能な、および/または転移性の疾患に対する少なくとも1つの先の全身療法または治療方針の際に進行していた食道がんである。一部の態様において、腫瘍はトリプルネガティブ乳がんである。一部の態様において、腫瘍は、不治の、進行型のエストロゲン受容体陰性、プロゲステロン受容体陰性、およびヒト上皮増殖因子受容体2陰性乳腺がんとして組織学的に確認されているトリプルネガティブ乳がんである。 In some embodiments, the invention inhibits tumor growth in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist (e.g., A method of treating cancer, wherein the tumor is high-frequency microsatellite-unstable colorectal cancer, microsatellite-stable colorectal cancer, triple-negative breast cancer, Merkel cell carcinoma, endometrial cancer, or esophageal cancer. It is cancer, provides a method. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises combining a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT with a PD-1 antagonist. wherein the tumor is microsatellite unstable colorectal cancer, microsatellite stable colorectal cancer, triple negative breast cancer, Merkel cell carcinoma, endometrial cancer, or esophageal cancer. and that bind to human TIGIT are heavy chain CDR1s containing TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises combining a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT with a PD-L1 antagonist. wherein the tumor is microsatellite unstable colorectal cancer, microsatellite stable colorectal cancer, triple negative breast cancer, Merkel cell carcinoma, endometrial cancer, or esophageal cancer. and that bind to human TIGIT are heavy chain CDR1s containing TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the tumor is esophageal cancer. In some embodiments, the tumor is esophageal cancer that has progressed during at least one prior systemic therapy or course of treatment for unresectable and/or metastatic disease. In some embodiments, the tumor is triple negative breast cancer. In some embodiments, the tumor is triple-negative breast cancer histologically confirmed as incurable, advanced estrogen receptor-negative, progesterone-receptor-negative, and human epidermal growth factor receptor 2-negative breast adenocarcinoma. be.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、対象において腫瘍成長を阻害する(例えば、がんを処置する)方法は、ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量をPD-L1アンタゴニストと組み合わせて対象に投与する工程を含み、腫瘍は高頻度MSI固形腫瘍であり、かつヒトTIGITに結合する抗体は、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、高頻度マイクロサテライト不安定性固形腫瘍には、MSI CRC、MSI胃がん、MSI子宮内膜がん、MSI卵巣がん、MSI肝胆道がん、MSI尿路がん、MSI脳がん、またはMSI皮膚がんが含まれ得る。 In some embodiments, the invention inhibits tumor growth in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist (e.g., treating cancer), wherein the tumor is a high-frequency MSI solid tumor. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises combining a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT with a PD-1 antagonist. wherein the tumor is a high-frequency MSI solid tumor, and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises combining a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT with a PD-L1 antagonist. wherein the tumor is a high-frequency MSI solid tumor, and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, high frequency microsatellite unstable solid tumors include MSI CRC, MSI gastric cancer, MSI endometrial cancer, MSI ovarian cancer, MSI hepatobiliary cancer, MSI urinary tract cancer, MSI brain. cancer, or MSI skin cancer.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、腫瘍は、抗PD-1抗体を用いる処置に対して抵抗性または不応性である。一部の態様において、対象において腫瘍成長を阻害する(例えば、がんを処置する)方法は、ヒトTIGITの細胞外ドメインに特異的に結合する治療的有効量の抗体を対象に投与する工程を含み、腫瘍は、PD-L1アンタゴニストを用いる処置に対して抵抗性または不応性であり、かつヒトTIGITに結合する抗体は、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、腫瘍は、抗PD-L1抗体を用いる処置に対して抵抗性または不応性である。 In some embodiments, the invention provides a method of inhibiting tumor growth (eg, treating cancer) in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent, wherein the tumor is is resistant or refractory to treatment with a PD-1 antagonist or a PD-L1 antagonist. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT. wherein the tumor is resistant or refractory to treatment with a PD-1 antagonist, and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the tumor is resistant or refractory to treatment with an anti-PD-1 antibody. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT. wherein the tumor is resistant or refractory to treatment with a PD-L1 antagonist, and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the tumor is resistant or refractory to treatment with an anti-PD-L1 antibody.

In some embodiments, the tumor that is resistant or refractory to treatment with a PD-1 antagonist or PD-L1 antagonist is melanoma, NSCLC, renal cell carcinoma, head and neck squamous cell carcinoma, urinary tract selected from the group consisting of cutaneous carcinoma, colorectal cancer (eg MSI or dMMR metastatic CRC), and hepatocellular carcinoma.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、PD-1アンタゴニストは抗PD-1抗体である。一部の態様において、抗PD-1抗体はペンブロリズマブ(キイトルーダ(KEYTRUDA))である。一部の態様において、抗PD-1抗体はニボルマブ(オプジーボ(OPDIVO))である。一部の態様において、対象において腫瘍成長を阻害する(がんを処置する)方法は、ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量をPD-L1アンタゴニストと組み合わせて対象に投与する工程を含み、腫瘍は、単一の作用物質としてPD-L1アンタゴニストを用いる処置に対して抵抗性または不応性であり、かつヒトTIGITに結合する抗体は、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、PD-L1アンタゴニストは抗PD-L1抗体である。一部の態様において、抗PD-L1抗体はアテゾリズマブ(テセントリク(TECENTRIQ))である。一部の態様において、抗PD-L1抗体はアベルマブ(MSB0010718C)である。 In some embodiments, the invention inhibits tumor growth in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist (e.g., treating cancer), wherein the tumor is resistant or refractory to treatment with a PD-1 antagonist as a single agent or a PD-L1 antagonist as a single agent. offer. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises combining a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT with a PD-1 antagonist. wherein the tumor is resistant or refractory to treatment with a PD-1 antagonist as a single agent, and the antibody that binds to human TIGIT is TSDYAWN (SEQ ID NO: :4) the heavy chain CDR1, including

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the PD-1 antagonist is an anti-PD-1 antibody. In some embodiments, the anti-PD-1 antibody is pembrolizumab (KEYTRUDA). In some embodiments, the anti-PD-1 antibody is nivolumab (OPDIVO). In some embodiments, a method of inhibiting tumor growth (treating cancer) in a subject comprises combining a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT with a PD-L1 antagonist. wherein the tumor is resistant or refractory to treatment with a PD-L1 antagonist as a single agent, and an antibody that binds to human TIGIT is TSDYAWN (SEQ ID NO:4 ), including the heavy chain CDR1,

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the PD-L1 antagonist is an anti-PD-L1 antibody. In some embodiments, the anti-PD-L1 antibody is atezolizumab (TECENTRIQ). In some embodiments, the anti-PD-L1 antibody is avelumab (MSB0010718C).

In some embodiments, the tumor is resistant or refractory to treatment with a PD-1 antagonist as a single agent or a PD-L1 antagonist as a single agent, melanoma, NSCLC, renal It may include cell carcinoma, head and neck squamous cell carcinoma, urothelial carcinoma, colorectal carcinoma (eg MSI or dMMR metastatic CRC), and hepatocellular carcinoma.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、対象において腫瘍成長を阻害する(例えば、がんを処置する)方法は、ヒトTIGITの細胞外ドメインに特異的に結合する治療的有効量の抗体を対象に投与する工程を含み、対象はPD-L1アンタゴニストを用いて以前に処置されたことがあり、かつヒトTIGITに結合する抗体は、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、対象は、単一の作用物質としてPD-1アンタゴニストまたはPD-L1アンタゴニストを用いて以前に処置されたことがある。一部の態様において、対象はPD-1アンタゴニストまたはPD-L1アンタゴニストを用いて以前に処置されたことがあり、処置中にまたは処置後に疾患進行があった。特定の処置の際に、または特定の処置の後に疾患進行があった対象は本明細書において「進行者」と呼ばれることがある。一部の態様において、対象は、第一選択治療薬である抗PD-1抗体を用いて以前に処置されたことがある。一部の態様において、対象は、第二選択治療薬または第三選択治療薬である抗PD-1抗体を用いて以前に処置されたことがある。一部の態様において、対象はペンブロリズマブ(キイトルーダ)を用いて以前に処置されたことがある。一部の態様において、対象はニボルマブ(オプジーボ)を用いて以前に処置されたことがある。一部の態様において、対象は、第一選択治療薬である抗PD-L1抗体を用いて以前に処置されたことがある。一部の態様において、対象は、第二選択治療薬または第三選択治療薬である抗PD-L1抗体を用いて以前に処置されたことがある。一部の態様において、対象はアテゾリズマブ(テセントリク)を用いて以前に処置されたことがある。一部の態様において、対象はアベルマブ(MSB0010718C)を用いて以前に処置されたことがある。一部の態様において、対象は、併用療法の一環としてPD-1アンタゴニストまたはPD-L1アンタゴニストを用いて以前に処置されたことがある。一部の態様において、対象は、抗PD-1抗体および第2のチェックポイント阻害物質を用いて以前に処置されたことがある。一部の態様において、対象は、抗PD-1抗体および抗CTLA-4抗体を用いて以前に処置されたことがある。一部の態様において、対象は、ニボルマブ(オプジーボ)およびイピリムマブ(ヤーボイ(YERVOY))を用いて以前に処置されたことがある。一部の態様において、対象は、抗PD-1抗体および化学療法剤を用いて以前に処置されたことがある。一部の態様において、対象は、抗PD-L1抗体および第2のチェックポイント阻害物質を用いて以前に処置されたことがある。一部の態様において、対象は、抗PD-L1抗体および抗CTLA-4抗体を用いて以前に処置されたことがある。一部の態様において、対象は、抗PD-L1抗体およびイピリムマブ(ヤーボイ)を用いて以前に処置されたことがある。一部の態様において、対象は抗PD-L1抗体および化学療法剤を用いて以前に処置されたことがある。 In some embodiments, the invention provides a method of inhibiting tumor growth (e.g., treating cancer) in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent, wherein the subject is A method is provided wherein the patient has been previously treated with a PD-1 antagonist or a PD-L1 antagonist. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT. wherein the subject has been previously treated with a PD-1 antagonist and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT. wherein the subject has been previously treated with a PD-L1 antagonist and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the subject has been previously treated with a PD-1 antagonist or a PD-L1 antagonist as a single agent. In some embodiments, the subject has been previously treated with a PD-1 antagonist or PD-L1 antagonist and has had disease progression during or after treatment. A subject who has had disease progression during or after a particular treatment may be referred to herein as a "progressor." In some embodiments, the subject has been previously treated with a first line therapeutic agent, an anti-PD-1 antibody. In some embodiments, the subject has been previously treated with an anti-PD-1 antibody that is a second-line or third-line therapy. In some embodiments, the subject has been previously treated with pembrolizumab (Keytruda). In some embodiments, the subject has been previously treated with nivolumab (Opdivo). In some embodiments, the subject has been previously treated with a first line therapeutic agent, an anti-PD-L1 antibody. In some embodiments, the subject has been previously treated with an anti-PD-L1 antibody that is a second-line or third-line therapy. In some embodiments, the subject has been previously treated with atezolizumab (Tecentriq). In some embodiments, the subject has been previously treated with avelumab (MSB0010718C). In some embodiments, the subject has been previously treated with a PD-1 antagonist or PD-L1 antagonist as part of combination therapy. In some embodiments, the subject has been previously treated with an anti-PD-1 antibody and a second checkpoint inhibitor. In some embodiments, the subject has been previously treated with an anti-PD-1 antibody and an anti-CTLA-4 antibody. In some embodiments, the subject has been previously treated with nivolumab (Opdivo) and ipilimumab (YERVOY). In some embodiments, the subject has been previously treated with an anti-PD-1 antibody and a chemotherapeutic agent. In some embodiments, the subject has been previously treated with an anti-PD-L1 antibody and a second checkpoint inhibitor. In some embodiments, the subject has been previously treated with an anti-PD-L1 antibody and an anti-CTLA-4 antibody. In some embodiments, the subject has been previously treated with an anti-PD-L1 antibody and ipilimumab (Yervoy). In some embodiments, the subject has been previously treated with an anti-PD-L1 antibody and a chemotherapeutic agent.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、対象は抗PD-1抗体を用いて以前に処置されたことがある。一部の態様において、対象はペンブロリズマブ(キイトルーダ)を用いて以前に処置されたことがある。一部の態様において、対象はニボルマブ(オプジーボ)を用いて以前に処置されたことがある。一部の態様において、抗PD-1抗体を用いる処置の間に腫瘍成長が進行した。一部の態様において、抗PD-1抗体を用いる処置の後に腫瘍成長が再発した。一部の態様において、腫瘍は、抗TIGIT抗体と抗PD-1抗体の併用療法に応答性である。一部の態様において、腫瘍は、抗TIGIT抗体と抗PD-1抗体の併用療法に応答性であり、この応答は、抗TIGIT抗体単独の投与を用いて得られる応答よりも優れている。一部の態様において、対象において腫瘍成長を阻害する(がんを処置する)方法は、ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量をPD-L1アンタゴニストと組み合わせて対象に投与する工程を含み、対象は、単一の作用物質としてPD-L1アンタゴニストを用いて以前に処置されたことがあり、かつヒトTIGITに結合する抗体は、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、対象は抗PD-L1抗体を用いて以前に処置されたことがある。一部の態様において、対象はアテゾリズマブ(テセントリク)を用いて以前に処置されたことがある。一部の態様において、対象はアベルマブ(MSB0010718C)を用いて以前に処置されたことがある。一部の態様において、抗PD-L1抗体を用いる処置の間に腫瘍成長が進行した。一部の態様において、抗PD-L1抗体を用いる処置の後に腫瘍成長が再発した。一部の態様において、腫瘍は、抗TIGIT抗体と抗PD-L1抗体の併用療法に応答性である。一部の態様において、腫瘍は、抗TIGIT抗体と抗PD-L1抗体の併用療法に応答性であり、この応答は、抗TIGIT抗体単独の投与を用いて得られる応答よりも優れている。 In some embodiments, the invention inhibits tumor growth (e.g., treats cancer) in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist. Methods are provided wherein the subject has been previously treated with a PD-1 antagonist or a PD-L1 antagonist. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises combining a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT with a PD-1 antagonist. wherein the subject has been previously treated with a PD-1 antagonist, and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the subject has been previously treated with an anti-PD-1 antibody. In some embodiments, the subject has been previously treated with pembrolizumab (Keytruda). In some embodiments, the subject has been previously treated with nivolumab (Opdivo). In some embodiments, tumor growth progressed during treatment with anti-PD-1 antibody. In some embodiments, tumor growth recurred after treatment with anti-PD-1 antibodies. In some embodiments, the tumor is responsive to combination therapy with an anti-TIGIT antibody and an anti-PD-1 antibody. In some embodiments, the tumor is responsive to combination therapy with an anti-TIGIT antibody and an anti-PD-1 antibody, and the response is superior to that obtained using administration of the anti-TIGIT antibody alone. In some embodiments, a method of inhibiting tumor growth (treating cancer) in a subject comprises combining a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT with a PD-L1 antagonist. wherein the subject has been previously treated with a PD-L1 antagonist as a single agent, and an antibody that binds human TIGIT is TSDYAWN (SEQ ID NO:4) heavy chain CDR1, including

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the subject has been previously treated with an anti-PD-L1 antibody. In some embodiments, the subject has been previously treated with atezolizumab (Tecentriq). In some embodiments, the subject has been previously treated with avelumab (MSB0010718C). In some embodiments, tumor growth progressed during treatment with anti-PD-L1 antibody. In some embodiments, tumor growth recurred after treatment with an anti-PD-L1 antibody. In some embodiments, the tumor is responsive to combination therapy with an anti-TIGIT antibody and an anti-PD-L1 antibody. In some embodiments, the tumor is responsive to combination therapy with an anti-TIGIT antibody and an anti-PD-L1 antibody, and the response is superior to that obtained using administration of the anti-TIGIT antibody alone.

In some embodiments, the tumor in a subject previously treated with a PD-1 antagonist or PD-L1 antagonist is a tumor such as melanoma, NSCLC, renal cell carcinoma, head and neck squamous cell carcinoma, May have urothelial carcinoma, colorectal cancer (eg, MSI or dMMR metastatic CRC), or hepatocellular carcinoma.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、腫瘍試料が対象から得られ、PVRおよび/またはPVRL2の発現レベルが決定される。一部の態様において、PVRの発現レベルは、処置のために対象を選択するのに用いられる。一部の態様において、PVRL2の発現レベルは、処置のために対象を選択するのに用いられる。 In some embodiments, the invention provides a method of inhibiting tumor growth (e.g., treating cancer) in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent, wherein the tumor/ A method is provided, wherein the cancer expresses PVR and/or PVRL2. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT. the tumor/cancer expresses PVR and/or PVRL2 and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, a tumor sample is obtained from the subject and the expression level of PVR and/or PVRL2 is determined. In some embodiments, PVR expression levels are used to select subjects for treatment. In some embodiments, the expression level of PVRL2 is used to select subjects for treatment.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments, the invention comprises selectively administering a therapeutically effective amount of a TIGIT binding agent to the subject based on the tumor/cancer expressing PVR and/or PVRL2. Methods of inhibiting tumor growth (eg, treating cancer) are provided. In some embodiments, a method of inhibiting tumor growth (e.g., treating cancer) in a subject is based on the tumor/cancer expressing PVR and/or PVRL2 to the extracellular domain of human TIGIT. selectively administering to the subject a therapeutically effective amount of an antibody that specifically binds, and wherein the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

, light chain CDR2 with SASYRYT (SEQ ID NO:8), and light chain CDR3 with QQHYSTP (SEQ ID NO:9).

In some embodiments, tumors/cancers expressing PVR and/or PVRL2 include NSCLC, renal cell carcinoma, head and neck squamous cell carcinoma, urothelial carcinoma, colorectal carcinoma (e.g., MSI or dMMR metastatic CRC), or hepatocellular carcinoma.

In some embodiments, the tumor/cancer expressing PVR and/or PVRL2 includes CRC, gastric cancer, endometrial cancer, ovarian cancer, hepatobiliary cancer, urinary tract cancer, brain cancer, or Includes skin cancer.

In some embodiments, tumors/cancers expressing PVR and/or PVRL2 include MSI CRC, MSI gastric cancer, MSI endometrial cancer, MSI ovarian cancer, MSI hepatobiliary cancer, MSI urinary tract cancer , MSI brain cancer, or MSI skin cancer.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、対象において腫瘍成長を阻害する(例えば、がんを処置する)方法は、ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量をPD-L1アンタゴニストと組み合わせて対象に投与する工程を含み、がんはPVRおよび/またはPVRL2を発現し、かつヒトTIGITに結合する抗体は、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、抗TIGIT抗体は抗PD-1抗体と組み合わせて投与される。一部の態様において、抗TIGIT抗体はペンブロリズマブ(キイトルーダ)と組み合わせて投与される。一部の態様において、抗TIGIT抗体はニボルマブ(オプジーボ)と組み合わせて投与される。一部の態様において、抗TIGIT抗体は抗PD-L1抗体と組み合わせて投与される。一部の態様において、抗TIGIT抗体はアテゾリズマブ(テセントリク)と組み合わせて投与される。 In some embodiments, the invention inhibits tumor growth in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist (e.g., A method of treating cancer is provided, wherein the cancer expresses PVR and/or PVRL2. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises combining a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT with a PD-1 antagonist. wherein the cancer expresses PVR and/or PVRL2 and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises combining a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT with a PD-L1 antagonist. wherein the cancer expresses PVR and/or PVRL2 and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, an anti-TIGIT antibody is administered in combination with an anti-PD-1 antibody. In some embodiments, an anti-TIGIT antibody is administered in combination with pembrolizumab (Keytruda). In some embodiments, an anti-TIGIT antibody is administered in combination with nivolumab (Opdivo). In some embodiments, an anti-TIGIT antibody is administered in combination with an anti-PD-L1 antibody. In some embodiments, an anti-TIGIT antibody is administered in combination with atezolizumab (Tecentriq).

In some embodiments, the expression level of PVR and/or PVRL2 in the tumor sample has been determined to be high. In some embodiments, the expression level of PVR in the sample is compared to a predetermined expression level of PVR. In some embodiments, the expression level of PVRL2 in the sample is compared to a predetermined expression level of PVRL2. In some embodiments, the predetermined expression level of PVR expression is the expression level of PVR in a reference tumor sample, a reference normal tissue sample, a series of reference tumor samples, or a series of reference normal tissue samples. In some embodiments, the predetermined expression level of PVRL2 expression is the expression level of PVRL2 in a reference tumor sample, a reference normal tissue sample, a series of reference tumor samples, or a series of reference normal tissue samples. In some embodiments, the expression level of PVR or PVRL2 is determined using an immunohistochemistry (IHC) assay. In some embodiments, the expression level of PVR and/or PVRL2 is determined using an assay comprising H-score evaluation. In some embodiments, the expression level of PVR is determined using an antibody that specifically binds PVR. In some embodiments, the expression level of PVRL2 is determined using an antibody that specifically binds PVRL2. In some embodiments, PVR is detected on the surface of tumor cells. In some embodiments, PVRL2 is detected on the surface of tumor cells. In some embodiments, PVR is detected on the surface of tumor-infiltrating immune cells and/or within the tumor microenvironment. In some embodiments, PVRL2 is detected on the surface of tumor-infiltrating immune cells and/or within the tumor microenvironment.

In certain embodiments, the expression level of PVR and/or PVRL2 is determined by reverse transcription PCR (RT-PCR), quantitative RT-PCR (qPCR), TaqMan™, or TaqMan™ Low Density Array (TLDA), etc. is determined using a PCR-based method, including but not limited to. In some embodiments, biomarker expression levels are determined using microarrays.

In certain embodiments, the expression level of PVR and/or PVRL2 is determined using protein-based methods. In some embodiments, the biomarker expression level is determined by multi-analyte profile test, radioimmunoassay (RIA), western blot assay, immunofluorescence assay, enzyme immunoassay, enzyme-linked immunosorbent assay (ELISA), Measured or determined by immunoprecipitation assay, chemiluminescence assay, immunohistochemical (IHC) assay, dot blot assay, or slot blot assay. In some embodiments, the expression level of PVR and/or PVRL2 is determined using an IHC assay. In some embodiments, the assay uses antibodies (eg, anti-PVR or anti-PVRL2 antibodies). In some embodiments where the assay uses an antibody, said antibody is detectably labeled. In some embodiments, the label is selected from the group consisting of immunofluorescent labels, chemiluminescent labels, phosphorescent labels, enzymatic labels, radiolabels, avidin/biotin labels, colloidal gold particles, colored particles, and magnetic particles. be done.

In some embodiments, PVR and/or PVRL2 expression levels are determined prior to treatment with a TIGIT binding agent. In some embodiments, a TIGIT binding agent is administered to a subject if the tumor or cancer has high expression levels of PVR and/or PVRL2. In some embodiments, the method comprises the steps of: (i) obtaining a tumor sample from the subject; (ii) measuring the expression level of PVR and/or PVRL2 in the sample; and (iii) tumor or cancer PVR and /or administering to the subject an effective amount of a TIGIT binding agent if the expression level of PVRL2 is elevated or high.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、対象において腫瘍成長を阻害する(例えば、がんを処置する)方法は、腫瘍のPVRおよび/またはPVRL2発現レベルが高いことに基づいて、ヒトTIGITの細胞外ドメインに特異的に結合する抗体の治療的有効量をPD-L1アンタゴニストと組み合わせて対象に選択的に投与する工程を含み、かつヒトTIGITに結合する抗体は、TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、抗TIGIT抗体は抗PD-1抗体と組み合わせて投与される。一部の態様において、抗TIGIT抗体はペンブロリズマブ(キイトルーダ)と組み合わせて投与される。一部の態様において、抗TIGIT抗体はニボルマブ(オプジーボ)と組み合わせて投与される。一部の態様において、抗TIGIT抗体は抗PD-L1抗体と組み合わせて投与される。一部の態様において、抗TIGIT抗体はアテゾリズマブ(テセントリク)と組み合わせて投与される。 In some embodiments, the present invention selects a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist for a subject based on high PVR and/or PVRL2 expression levels in the tumor. A method of inhibiting tumor growth (eg, treating cancer) in a subject is provided, comprising the step of administering targetally. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises combining a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT with a PD-1 antagonist. wherein the antibody that has high tumor PVR and/or PVRL2 expression levels and that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, methods of inhibiting tumor growth (e.g., treating cancer) in a subject are specific for the extracellular domain of human TIGIT based on high PVR and/or PVRL2 expression levels in tumors. selectively administering to the subject a therapeutically effective amount of an antibody that binds to a PD-L1 antagonist in combination with a PD-L1 antagonist, and wherein the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4) ,

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, an anti-TIGIT antibody is administered in combination with an anti-PD-1 antibody. In some embodiments, an anti-TIGIT antibody is administered in combination with pembrolizumab (Keytruda). In some embodiments, an anti-TIGIT antibody is administered in combination with nivolumab (Opdivo). In some embodiments, an anti-TIGIT antibody is administered in combination with an anti-PD-L1 antibody. In some embodiments, an anti-TIGIT antibody is administered in combination with atezolizumab (Tecentriq).

In some embodiments, the sample is a biopsy sample. In some embodiments, the sample comprises tumor cells, cancer-infiltrating immune cells, stromal cells, and any combination thereof. In some embodiments, the sample is a formalin-fixed paraffin-embedded (FFPE) sample. In some embodiments, the sample is archival tissue, fresh tissue, or frozen tissue.

Some patients have tumors with an active immune microenvironment. These tumors are termed 'inflammatory' or 'hot' tumors and are characterized, for example, by the presence of tumor-infiltrating lymphocytes (TILs), IFN-γ-producing CD8+ T cells, and PD-L1 expression. In several studies, inflammatory or hot tumors have been associated with clinical responses to immunotherapy.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、腫瘍は、予め決定されたレベルと比較して高レベルのTILを含む。一部の態様において、TILのレベルはTILの絶対数である。一部の態様において、TILのレベルは、総細胞数に対するパーセントである。 Accordingly, in some aspects, the invention provides a method of inhibiting tumor growth (e.g., treating cancer) in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent, comprising: A method is provided, wherein the tumor comprises TILs. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT. the tumor comprises TILs, and the antibody that binds to human TIGIT comprises a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

, light chain CDR2 with SASYRYT (SEQ ID NO:8), and light chain CDR3 with QQHYSTP (SEQ ID NO:9). In some embodiments, the tumor contains elevated levels of TILs compared to a predetermined level. In some embodiments, the level of TILs is the absolute number of TILs. In some embodiments, the level of TILs is a percentage of total cell number.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、腫瘍は、予め決定されたレベルと比較して高レベルのTILを含む。一部の態様において、TILのレベルはTILの絶対数である。一部の態様において、TILのレベルは、総細胞数に対するパーセントである。一部の態様において、抗TIGIT抗体は抗PD-1抗体と組み合わせて投与される。一部の態様において、抗TIGIT抗体はペンブロリズマブ(キイトルーダ)と組み合わせて投与される。一部の態様において、抗TIGIT抗体はニボルマブ(オプジーボ)と組み合わせて投与される。一部の態様において、抗TIGIT抗体は抗PD-L1抗体と組み合わせて投与される。一部の態様において、抗TIGIT抗体はアテゾリズマブ(テセントリク)と組み合わせて投与される。 In some embodiments, the invention inhibits tumor growth in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist (e.g., treatment of cancer), wherein the tumor comprises TILs. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject includes administering a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT as a PD-1 antagonist or PD. - administering to the subject in combination with an L1 antagonist, wherein the tumor comprises TILs, and the antibody that binds to human TIGIT comprises a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the tumor contains elevated levels of TILs compared to a predetermined level. In some embodiments, the level of TILs is the absolute number of TILs. In some embodiments, the level of TILs is a percentage of total cell number. In some embodiments, an anti-TIGIT antibody is administered in combination with an anti-PD-1 antibody. In some embodiments, an anti-TIGIT antibody is administered in combination with pembrolizumab (Keytruda). In some embodiments, an anti-TIGIT antibody is administered in combination with nivolumab (Opdivo). In some embodiments, an anti-TIGIT antibody is administered in combination with an anti-PD-L1 antibody. In some embodiments, an anti-TIGIT antibody is administered in combination with atezolizumab (Tecentriq).

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:7)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:8)を含む軽鎖CDR3を含む。 In some embodiments, the invention provides a method of inhibiting tumor growth (eg, treating cancer) in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent, wherein the tumor is Methods are provided comprising high levels of regulatory T cells (Treg). In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT. the tumor contains high levels of Tregs, and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:7), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:8).

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、抗TIGIT抗体は抗PD-1抗体と組み合わせて投与される。一部の態様において、抗TIGIT抗体はペンブロリズマブ(キイトルーダ)と組み合わせて投与される。一部の態様において、抗TIGIT抗体はニボルマブ(オプジーボ)と組み合わせて投与される。一部の態様において、抗TIGIT抗体は抗PD-L1抗体と組み合わせて投与される。一部の態様において、抗TIGIT抗体はアテゾリズマブ(テセントリク)と組み合わせて投与される。 In some embodiments, the invention inhibits tumor growth in a subject comprising administering to the subject a therapeutically effective amount of a TIGIT binding agent in combination with a PD-1 antagonist or PD-L1 antagonist (e.g., treatment of cancer), wherein the tumor contains high levels of Tregs. In some embodiments, a method of inhibiting tumor growth (eg, treating cancer) in a subject includes administering a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of human TIGIT as a PD-1 antagonist or PD. - administering to the subject in combination with an L1 antagonist, wherein the tumor contains high levels of Tregs, and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, an anti-TIGIT antibody is administered in combination with an anti-PD-1 antibody. In some embodiments, an anti-TIGIT antibody is administered in combination with pembrolizumab (Keytruda). In some embodiments, an anti-TIGIT antibody is administered in combination with nivolumab (Opdivo). In some embodiments, an anti-TIGIT antibody is administered in combination with an anti-PD-L1 antibody. In some embodiments, an anti-TIGIT antibody is administered in combination with atezolizumab (Tecentriq).

In some embodiments, the presence of immune cells (e.g., TILs, Tregs, Teff) in a tumor or within the tumor microenvironment is determined by assessing the expression of specific proteins on the cell surface or intracellularly. analyzed. Commonly used methods for analyzing protein expression include, but are not limited to, immunohistochemistry (IHC)-based methods, antibody-based methods, and mass spectrometry-based methods. In some embodiments, protein expression on the surface of immune cells or within immune cells is measured by multiple analyte profile test, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay, western blot assay, immunofluorescence assay, enzyme immunoassay. , immunoprecipitation assays, chemiluminescence assays, immunohistochemical (IHC) assays, dot blot assays, or slot blot assays. Antibodies, typically monoclonal antibodies, may be used to detect expression of gene products (eg, proteins). In some embodiments, the antibody can be detected by directly labeling the antibody itself. In other embodiments, an unlabeled primary antibody is used together with a labeled secondary antibody.

In some embodiments, expression of a particular protein is measured using an agent that specifically binds to that protein. Any molecular entity that exhibits specific binding to that protein can be used to assess protein expression in a sample. Specific binding agents include, but are not limited to, antibodies, antibody mimetics, and polynucleotides (eg, aptamers). Those skilled in the art will appreciate that the degree of specificity required will depend on the particular assay used to detect the protein.

CD45 (leukocyte common antigen, LCA; includes all proteins in the CD45 family) is used as a marker for immune cells including lymphocytes, NK cells, macrophages, monocytes and the like. T cells can be separated into different subsets including but not limited to CD4+ T cells or CD8+ T cells. In some embodiments, CD45 is used as a TIL marker in tumor samples. In some embodiments, TILs are detected and/or assessed using anti-CD45 antibodies. In some embodiments, a cocktail of monoclonal anti-CD45 antibodies is used, each recognizing a different CD45 isotype and/or a different epitope. In some embodiments, TILs are detected and/or assessed using anti-CD4 antibodies. In some embodiments, TILs are detected and/or assessed using anti-CD8 antibodies.

It is known to those skilled in the art that FOXP3 is a regulatory T cell (Treg) marker and can be used to detect and/or identify Tregs in tumor samples. Studies have shown that Treg cells are composed of many subpopulations with different functions, and one subpopulation has been identified as FOXP3+ and TIGIT+. In some embodiments, Tregs are detected and/or assessed by FOXP3 expression. In some embodiments, FOXP3 expression is detected and/or assessed using an anti-FOXP3 antibody. In some embodiments, a cocktail of monoclonal anti-FOXP3 antibodies are used, each recognizing a different isotype and/or a different epitope. In some embodiments, Tregs are detected and/or assessed by FOXP3 and TIGIT expression. In some embodiments, TIGIT expression is detected and/or assessed using an anti-TIGIT antibody.

In some aspects, an antibody is used in the assay, and the antibody is detectably labeled. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, phosphorescent materials, luminescent materials, bioluminescent materials, chemiluminescent materials, radioactive materials, avidin/biotin, colloidal gold particles, colored particles, and magnetic particles. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, ?-galactosidase, or acetylcholinesterase. Examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin. Examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride, or phycoerythrin. An example of a luminescent material includes luminol. Examples of bioluminescent materials include luciferase, luciferin, and aequorin. Examples of suitable radioactive materials include ¹²⁵ I, ¹³¹ I, ³⁵ S, or ³ H.

In some embodiments, CD45, CD4, CD8, FOXP3, and/or TIGIT expression is measured by an IHC assay. For example, FFPE sections are cut from tumor samples and mounted on coated glass slides. Deparaffinize and rehydrate the tissue by sequentially incubating the tissue in xylene, 100% ethanol, 95% ethanol, 70% ethanol, and distilled water for antigen retrieval. Slides are placed in retrieval solution and placed in a decloaker for antigen retrieval. To block endogenous peroxidase activity, slides are incubated in hydrogen peroxide and washed with PBS. Slides are incubated in blocker to block non-specific background staining. Incubate slides with anti-PD-L1 antibody. Specific binding is detected using a kit containing diaminobenzidine (DAB). Sections are counterstained with hematoxylin. In some embodiments, FFPE sections are mounted on coated glass slides and stained using an automated system, eg, a Ventana BenchMark ULTRA instrument using Ventana reagents.

In some embodiments, said antibody is a polyclonal antibody. In some embodiments, said antibody is a monoclonal antibody. In some embodiments, the antibody is a murine antibody that binds to human biomarkers (eg, CD45, CD4, CD8, FOXP3, TIGIT). In some embodiments, the antibody is a rabbit antibody that binds to human biomarkers (eg, CD45, CD4, CD8, FOXP3, TIGIT). In some embodiments, the antibody is a goat antibody that binds to human biomarkers (eg, CD45, CD4, CD8, FOXP3, TIGIT).

The IHC slides may be analyzed using automated instruments or manually evaluated microscopically. The staining intensity of each cell (0: no expression, 1: weak expression, 2: intermediate expression, 3: strong expression) is measured, cells with each staining level are counted and the percentage of each type is calculated. Data are summarized for each tissue section into a weighted H-score: H-score = [3 x (3 + cell %)] + [2 x (2 + cell %)] + [1 x (1 + cell %)]. With these parameters, the highest usable score is H-score=300. In some embodiments, an H-score of 1 or less is considered negative. In some embodiments, the IHC assay has a cutoff value. In some embodiments, the IHC assay has a specificity cutoff value. In some embodiments, the IHC assay has a cutoff value for efficacy. In some embodiments, the IHC assay has a cutoff value determined by screening positive and negative tumor tissue. In some embodiments, the IHC assay has a cutoff value of about 25. In some embodiments, the IHC assay cutoff value is about 30, about 40, about 50, about 60, about 70, about 80, about 90, about 100, or about 110, or about 120.

Other suitable methods for analyzing biomarker expression levels include proteomics-based methods. Proteomics specifically includes the study of global protein expression changes in samples. In some embodiments, the proteomic method comprises the following steps: (1) separation of individual proteins in the sample by 2D electrophoresis (2-D PAGE), (2) (e.g., mass spectrometry or N-terminal identification of individual proteins recovered from the gel (by sequencing) and (3) analysis of the data using bioinformatics. In some embodiments, proteomic methods involve the use of tissue microarrays (TMA). Tissue arrays can be constructed according to various techniques known to those of skill in the art. In certain embodiments, a manual tissue arrayer is used to remove "cores" from paraffin blocks prepared from tissue samples. The cores are then inserted at designated locations on the grid of another paraffin block. Cores from as many as 400 samples can be inserted into one recipient block. The resulting tissue array may be sectioned for analysis. In some embodiments, the proteomic method comprises an antibody microarray. In some embodiments, proteomic methods include the use of mass spectrometry methods, including but not limited to SELDI, MALDI, electrospray, and surface plasmon resonance methods. In some embodiments, proteomic methods include bead-based technologies including, but not limited to, antibodies on beads in an array format. In some embodiments, the proteomic method comprises reversed-phase protein microarrays (RPPM). In some embodiments, proteomic methods include multiplexed protein profiling, including but not limited to Global Proteome Survey (GPS) methods.

Other suitable methods for analyzing biomarker expression levels include, but are not limited to, methods based on analysis of polynucleotide expression, sequencing of polynucleotides, and/or analysis of protein expression. For example, determination of biomarker expression levels may be performed by detecting expression of mRNA expressed from the gene of interest and/or by detecting expression of the polypeptide encoded by that gene.

Commonly used methods for analyzing polynucleotides include Southern blot analysis, Northern blot analysis, and in situ hybridization, RNase protection assays, and polymerase chain reaction (PCR)-based methods, such as reverse transcription polymerase chain reaction. reaction (RT-PCR), quantitative PCR, also known as real-time PCR (qPCR), TaqMan™, TaqMan™ Low Density Array (TLDA), anchored PCR, competitive PCR, rapid amplification of cDNA ends (RACE) , and microarray analysis. RT-PCR is a quantitative method that can be used to examine gene expression profiles by comparing mRNA levels in different samples. A variation of RT-PCR is real-time quantitative PCR, which measures PCR product accumulation with dual-labeled fluorescent probes (eg, TaqMan™ probes). Many others known to those of skill in the art including, but not limited to, differential display, amplified fragment length polymorphism, BeadArray™ technology, high coverage expression profiling (HiCEP), and digital PCR. There are PCR-based techniques for Exemplary methods for sequencing-based gene expression analysis include Serial Analysis of Gene Expression (SAGE), Massively Parallel Signature Sequencing (MPSS), and NexGen sequencing, including mRNA sequencing. Includes analytics.

In certain embodiments, biomarker expression is measured using a qPCR assay. For example, extract total RNA from fresh-frozen (FF) tissue samples, or extract total RNA from macro-dissected formalin-fixed paraffin-embedded (FFPE) tissue samples. Quantity and quality of total RNA are assessed by standard spectrophotometry and/or any other suitable method (eg Agilent Bioanalyzer). After RNA extraction, RNA samples are reverse transcribed using standard methods and/or commercially available cDNA synthesis kits (eg, Roche Transcriptor First Strand cDNA Synthesis Kit). The resulting cDNA is pre-amplified using, for example, an ABI pre-amplification kit. Expression of biomarkers (eg, CD45, CD4, CD8, Foxp3, TIGIT, other immune response genes) is assessed using ABI TaqMan Gene Expression Mastermix, eg, by Roche Lightcycler 480 system (Roche Diagnostics). qPCR reactions are performed in triplicate. For each assay, a portion of the sample is run without reverse transcription (RT-negative control) and a control sample is run without template. A universal human reference RNA sample is included on each plate to serve as a positive control. Identify a suitable reference gene from a standard reference gene panel. To eliminate the risk of co-regulation, candidate reference genes with different cellular functions are selected. Specific software and algorithms (eg Genex software; GeNorm and Normfinder algorithms) are used to evaluate and select the most suitable reference genes. Expression levels of each biomarker are normalized using the best reference gene of choice. In some embodiments, these normalized (or standardized) expression values for each biomarker are used to calculate the determined value for the sample. In some embodiments, these normalized (or normalized) expression values for each biomarker are used to calculate expression levels.

In some embodiments, biomarker expression is measured using PCR-based assays comprising primers and/or probes specific for human biomarkers (eg, CD45, CD4, CD8, Foxp3, TIGIT). As used herein, the term "probe" refers to any molecule capable of selectively binding to a specifically intended target biomolecule. Probes may be synthesized by one skilled in the art using known techniques, or may be derived from biological preparations. Probes can include, but are not limited to, RNA, DNA, proteins, peptides, aptamers, antibodies, and organic molecules. The term "primer" or "probe" includes an oligonucleotide having a sequence of a particular SEQ ID NO or an oligonucleotide having a sequence complementary to a particular SEQ ID NO. In some embodiments, probes are modified. In some embodiments, the probe is modified with a quencher. In some embodiments, probes are labeled. Labels can include, but are not limited to, colorimetric, fluorescent, chemiluminescent, or bioluminescent labels.

Alternatively, biomarker expression levels may be determined by amplifying complementary DNA (cDNA) or complementary RNA (cRNA) generated from mRNA and analyzing using microarrays. Microarray technology makes it possible to analyze the expression of thousands of genes simultaneously. Many different array configurations and methods of generating array configurations are known to those skilled in the art. Additionally, microarrays are commercially available (eg, Affymetrix GeneChips) or can be custom made. Currently, widely used microarrays include cDNA arrays and oligonucleotide arrays. Generally, polynucleotides of interest (eg, probes or probe sets) are plated or arrayed on a microchip substrate. In some embodiments, probes for at least 10, 25, 50, 100, 500, 1000, 5000, 10,000, 20,000, or 25,000 or more genes are immobilized on the array substrate. The substrate can be a porous or non-porous support, such as a glass, plastic, or gel surface. Probes can include DNA, RNA, copolymer sequences of DNA and RNA, DNA analogs and/or RNA analogs, or combinations thereof. In some embodiments, the microarray comprises a support having an ordered array of binding sites for each gene. A microarray may be an addressable array or a positionally addressable array, e.g., each probe of the array may be attached to a solid support such that the identity of each probe is apparent from its position on the array. located at a known, predetermined position on the top.

Each probe on the microarray can be from 10 to 50,000 nucleotides in length. In some embodiments, the microarray probes are less than about 1,000 nucleotides in length, less than about 750 nucleotides, less than about 500 nucleotides, less than about 250 nucleotides, less than about 100 nucleotides, or less than about 50 nucleotides in length. It may consist of an array. Generally, the array contains positive control probes and negative control probes.

In certain embodiments, biomarker expression is measured using a microarray. For example, extract total RNA from fresh frozen (FF) tissue samples, or extract total RNA from formalin-fixed paraffin-embedded (FFPE) tissue samples obtained using microdissection. Total RNA quantity and quality is assessed by standard spectrophotometric methods and/or any other suitable technique (eg Agilent Bioanalyzer). After RNA extraction, RNA samples are amplified using standard methods and/or commercially available amplification systems (eg, NuGEN Ovation RNA Amplification System V2). Amplified cDNA is fragmented, labeled, and hybridized to microarrays (eg, using NuGEN Encore Biotin Modules and Affymetrix GeneChip arrays) according to standard procedures. Arrays are washed, stained and scanned according to the microarray instructions. Microarray data are preprocessed and probe-level intensity measurements are background-corrected, normalized using the Robust Multichip algorithm (RMA), and summarized as expression measurements. Combine probe level data to obtain expression levels for each biomarker (eg, CD45, CD4, CD8, Foxp3, TIGIT). A combination of quality parameter thresholds and data reduction methods (eg, principal component analysis) are applied to the dataset to establish profile quality and identify potential outlying samples. These normalized (or normalized) expression values for each biomarker are used to calculate the determined value for the sample.

In some embodiments of any of the methods described herein, the tumor/cancer is lung, liver, breast, renal cell carcinoma/kidney, prostate, gastrointestinal/stomach, melanoma, neck, bladder, selected from the group consisting of glioblastoma, head and neck, pancreatic, ovarian, colorectal, endometrial, and esophageal tumors/cancers. In some embodiments, the colorectal cancer is high frequency microsatellite unstable colorectal cancer. In some embodiments, the colorectal cancer is microsatellite stable colorectal cancer. In some embodiments, the esophageal tumor is histologically confirmed unresectable, advanced, or recurrent esophageal or gastroesophageal junction cancer. In some embodiments, the esophageal tumor is a tumor that has progressed during at least one prior systemic therapy or course of treatment for unresectable or metastatic disease. In some embodiments, the colorectal tumor is a histologically confirmed, incurable, advanced adenocarcinoma of the colon or rectum. In some embodiments, the colorectal tumor has been treated with at least one prior standard chemotherapy regimen for colorectal cancer or metastatic colorectal cancer and is refractory to these therapies or tumors that are progressing on these therapies. In some embodiments, the colorectal tumor has been treated with at least two prior standard chemotherapy regimens for colorectal cancer or metastatic colorectal cancer and is refractory to these therapies or tumors that are progressing at the time of these therapies. In some embodiments, the liver tumor or hepatocellular tumor is histologically confirmed advanced hepatocellular carcinoma that is not amenable to surgical and/or locoregional therapy. In some embodiments, the liver tumor or hepatocellular tumor has progressed or has progressed following surgical and/or locoregional therapy. In some embodiments, the cervical tumor is histologically confirmed recurrent or metastatic cervical cancer. In some embodiments, the cervical tumor is a tumor that has progressed during at least one prior course of chemotherapy for cervical cancer. In some embodiments, the breast tumor is triple negative breast cancer. In some embodiments, the breast tumor is histologically confirmed as incurable, advanced estrogen receptor-negative, progesterone receptor-negative, and human epidermal growth factor receptor 2-negative breast adenocarcinoma. In some embodiments, the head and neck tumor is histologically confirmed recurrent or metastatic squamous cell carcinoma of the head and neck (SCCHN) for which standard definitive or symptomatic therapy is not amenable. In some embodiments, head and neck tumors include, but are not limited to, oral cavity, nasal cavity, sinus, nasopharyngeal, oropharyngeal, hypopharyngeal, or laryngeal squamous cell carcinoma. In some embodiments, head and neck tumors include primary or recurrent cancers for which definitive therapy or established symptomatic therapy is not amenable.

In some embodiments of the methods described herein, the invention provides use of a TIGIT binding agent in the manufacture or preparation of a medicament for inhibiting growth of a tumor or tumor cells. In some aspects of the methods described herein, the invention provides use of a TIGIT binding agent in the manufacture or preparation of a medicament for treating cancer. In some embodiments, the TIGIT binding agent binds to human TIGIT and inhibits or reduces cancer growth. In certain embodiments, the tumor/cancer comprises cancer stem cells. In certain embodiments, the frequency of cancer stem cells in the tumor/cancer is reduced. In certain embodiments, the subject is human. In certain embodiments, the subject is at least partially depleted of a tumor.

Combination therapy with two or more therapeutic agents often employs agents that work by different mechanisms of action, but this is not required. Combination therapy using agents with different mechanisms of action may result in additive or synergistic effects. With combination therapy, the doses of each agent are lower than those used in monotherapy, thereby potentially reducing toxic side effects of the agents and/or increasing the therapeutic index of the agents. there is a possibility. Combination therapy may reduce the likelihood that resistant cancer cells will develop. In some embodiments, the combination therapy includes a therapeutic agent that affects the immune response (e.g., enhances or activates the response) and a tumor/cancer cell that affects (inhibits or inhibits the tumor/cancer cell). kill) including therapeutic substances.

In some embodiments of the methods described herein, combining the TIGIT binding agent and at least one additional therapeutic agent produces additive or synergistic results. In some embodiments, combination therapy increases the therapeutic index of the TIGIT binding agent. In some embodiments, combination therapy increases the therapeutic index of the additional therapeutic agent. In some embodiments, combination therapy reduces toxicity and/or side effects of the TIGIT binding agent. In some embodiments, combination therapy reduces toxicity and/or side effects of additional therapeutic agents.

In certain embodiments of the methods described herein, the method or treatment, in addition to administering a TIGIT binding agent, further comprises administering at least one additional therapeutic agent. The additional therapeutic agent can be administered before the agent is administered, at the same time as the agent is administered, and/or after the agent is administered. In some embodiments, the at least one additional therapeutic agent comprises 1, 2, 3, or more additional therapeutic agents.

Therapeutic agents that can be administered in combination with a TIGIT binding agent include chemotherapeutic agents. Thus, in some embodiments, the methods or treatments involve administering a TIGIT binding agent and a chemotherapeutic agent in combination or in combination with a cocktail of chemotherapeutic agents. Treatment with a TIGIT binding agent can occur before chemotherapy is administered, at the same time that chemotherapy is administered, or after chemotherapy is administered. Co-administration may involve co-administration in a single pharmaceutical formulation or co-administration using separate formulations, in either order, so that all of the active agents can exert their biological activity at the same time. It may also include continuous administration, generally within a period of time. Preparation and dosing schedules for such chemotherapeutic agents can be used according to manufacturers' instructions or as determined empirically by the skilled practitioner. Preparation and dosing regimens for such chemotherapy are also described in The Chemotherapy Source Book, 4th ^Edition , 2008, MC Perry, Editor, Lippincott, Williams & Wilkins, Philadelphia, PA.

Useful classes of therapeutic agents include, for example, antitubulin agents, auristatins, DNA minor groove binders, DNA replication inhibitors, alkylating agents (e.g., platinum complexes such as cisplatin, mono (platinum), bis (platinum), and trinuclear platinum complexes, and carboplatin), anthracyclines, antibiotics, antifolates, antimetabolites, chemotherapy sensitizers, duocarmycins, etoposide, fluoropyrimidines, ionophores, lexitropsin (lexitropsin), nitrosoureas, platinol, purine antimetabolites, puromycin, radiosensitizers, steroids, taxanes, topoisomerase inhibitors, vinca alkaloids, and the like. In certain embodiments, the second therapeutic agent is an alkylating agent, an antimetabolite, an antimitotic agent, a topoisomerase inhibitor, or an angiogenesis inhibitor.

Chemotherapeutic agents useful in the present invention include alkylating agents such as thiotepa and cyclosphosphamide (CYTOXAN); alkyl sulfonates such as busulfan, improsulfan, and piposulfan; aziridines such as benzodopa, carbocones, meturedopa, and uredopa; including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphoramide, and trimethylolomelamime; ethyleneimine and methylamelamine; nitrogen mustards such as chlorambucil, chlornafadine, cyclophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin ), phenesterine, prednimustine, trophosphamide, uracil mustard; nitrosoureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as aclacinomycin, actinomycin, authramycin, azaserine , bleomycin, cactinomycin, calicheamicin, carabicin, caminomycin, cardinophylline, chromomycin, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, ethorubicin, idarubicin, marcellomycin, mitomycin, mycophenolic acid, nogaramycin, olibomycin, peplomycin, potfiromycin, puromycin, quelamycin, rhodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, dinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, lopterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytosine arabinoside, dideoxyuridine, doxifluridine, enocitabine, floxuridine, 5-FU; androgens, such as carsterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; anti-adrenal agents, such as aminoglutethimide, mitotane, trilostane; acegratone; aldophosphamide glycoside; aminolevulinic acid; amsacrine; bestrabcil; bisantrene; edatraxate; elliptinium acetate; etogluside; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamole; nitracrine; pentostatin; spirogermanium; tenuazonic acid; triaziquone; 2,2',2''-trichlorotriethylamine; urethane; vindesine; dacarbazine; , paclitaxel (TAXOL) and docetaxel (TAXOTERE); chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; platinum analogues such as cisplatin and carboplatin; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbine; novantrone; teniposide; daunomycin; aminopterin; ibandronic acid; XELODA)); and pharmaceutically acceptable salts, acids, or derivatives of any of the foregoing. Chemotherapeutic agents include antihormonal agents that act to modulate or inhibit hormone action on tumors, e.g., tamoxifen, raloxifene, aromatase inhibitor 4(5)-imidazole, 4-hydroxytamoxifene, trioxifene, keoxifene , LY117018, onapristone, and toremifene (FARESTON), and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin, and pharmaceutically acceptable salts, acids of any of the foregoing. , or derivatives. In certain embodiments, the additional therapeutic agent is cisplatin. In certain embodiments, the additional therapeutic agent is carboplatin.

In certain embodiments, the chemotherapeutic agent is a topoisomerase inhibitor. Topoisomerase inhibitors are chemotherapeutic agents that interfere with the action of the topoisomerase enzyme (topoisomerase I or II). Topoisomerase inhibitors include doxorubicin HCl, daunorubicin citrate, mitoxantrone HCl, actinomycin D, etoposide, topotecan HCl, teniposide (VM-26), and irinotecan, and pharmaceutically acceptable salts of any of these , acids, or derivatives. In some embodiments, the additional therapeutic agent is irinotecan.

In certain embodiments, the chemotherapeutic agent is an antimetabolite. Antimetabolites are chemicals whose structures resemble metabolites required for normal biochemical reactions, but which differ sufficiently in structure to interfere with one or more normal functions of cells, such as cell division. Antimetabolites include gemcitabine, fluorouracil, capecitabine, methotrexate sodium, lalitrexed, pemetrexed, tegafur, cytosine arabinoside, thioguanine, 5-azacytidine, 6-mercaptopurine, azathioprine, 6-thioguanine, pentostatin, phosphorus Including, but not limited to, the acids fludarabine, and cladribine, and pharmaceutically acceptable salts, acids, or derivatives of any of these. In certain embodiments, the additional therapeutic agent is gemcitabine.

In certain embodiments, the chemotherapeutic agent is an antimitotic agent, including, but not limited to, agents that bind tubulin. In some embodiments, the agent is a taxane. In certain embodiments, the agent is paclitaxel or docetaxel, or a pharmaceutically acceptable salt, acid, or derivative of paclitaxel or docetaxel. In certain embodiments, the agent is paclitaxel (Taxol), docetaxel (Taxotere), albumin-bound paclitaxel (ABRAXANE®), DHA-paclitaxel, or PG-paclitaxel. In certain other aspects, the antimitotic agent comprises a vinca alkaloid, eg, vincristine, vinblastine, vinorelbine, or vindesine, or a pharmaceutically acceptable salt, acid, or derivative thereof. In some aspects, the antimitotic agent is an inhibitor of kinesin Eg5 or an inhibitor of mitotic kinases, eg, AuroraA or Plk1. In certain aspects, the additional therapeutic agent is paclitaxel. In certain embodiments, the additional therapeutic agent is albumin-bound paclitaxel (ABRAXANE®).

In some embodiments, additional therapeutic agents include agents such as small molecules. For example, treatment may involve co-administration of agents of the invention with small molecules that act as inhibitors against tumor-associated antigens, including but not limited to EGFR, HER2 (ErbB2), and/or VEGF. . In some embodiments, the agents of the invention are gefitinib (IRESSA), erlotinib (TARCEVA), sunitinib (SUTENT), lapatanib, vandetanib (ZACTIMA), Co-administered with a protein kinase inhibitor selected from the group consisting of AEE788, CI-1033, cediranib (RECENTIN), sorafenib (NEXAVAR), and pazopanib (GW786034B). In some embodiments, the additional therapeutic agent comprises an mTOR inhibitor.

In certain embodiments, the additional therapeutic agent is an agent that inhibits cancer stem cell pathways. In some embodiments, the additional therapeutic agent is an inhibitor of the Notch pathway. In some embodiments, the additional therapeutic agent is a Wnt pathway inhibitor. In some embodiments, the additional therapeutic agent is an inhibitor of the BMP pathway. In some embodiments, the additional therapeutic agent is an inhibitor of the Hippo pathway. In some embodiments, the additional therapeutic agent is an inhibitor of the RSPO/LGR pathway. In some embodiments, the additional therapeutic agent is an inhibitor of the mTOR/AKR pathway.

In some embodiments of the methods described herein, the additional therapeutic agent comprises a biological molecule such as an antibody. For example, treatment may involve co-administration of a TIGIT binding agent and an antibody against a tumor-associated antigen, including, but not limited to, an antibody that binds EGFR, HER2/ErbB2, and/or VEGF. In certain embodiments, the additional therapeutic agent is an antibody specific for a cancer stem cell marker. In some embodiments, the additional therapeutic agent is an antibody that binds to a component of the Notch pathway. In some embodiments, the additional therapeutic agent is an antibody that binds to a component of the Wnt pathway. In certain embodiments, the additional therapeutic agent is an antibody that inhibits cancer stem cell pathways. In some embodiments, the additional therapeutic agent is an inhibitor of the Notch pathway. In some embodiments, the additional therapeutic agent is a Wnt pathway inhibitor. In some embodiments, the additional therapeutic agent is an inhibitor of the BMP pathway. In some embodiments, the additional therapeutic agent is an antibody that inhibits β-catenin signaling. In certain embodiments, the additional therapeutic agent is an antibody that is an angiogenesis inhibitor (eg, an anti-VEGF antibody or an anti-VEGF receptor antibody). In certain embodiments, the additional therapeutic agent is bevacizumab (AVASTIN), ramucirumab, trastuzumab (HERCEPTIN), pertuzumab (OMNITARG), panitumumab (VECTIBIX), nimotuzumab (nimotuzumab), zartumumab, or cetuximab (ERBITUX).

In certain embodiments of the methods described herein, the method or treatment further comprises, in addition to administering a TIGIT binding agent, administering at least one additional immunotherapeutic agent. In some embodiments, the additional immunotherapeutic agent is an immune response stimulating agent. In some embodiments, immunotherapeutic agents (e.g., immune response stimulating agents) include colony-stimulating factors (e.g., granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), granulocyte colony stimulating factor (G-CSF), stem cell factor (SCF)), interleukins (e.g. IL-1, IL2, IL-3, IL-7, IL-12, IL-15, IL-18), Antibodies that block immunosuppressive function (e.g., anti-CTLA4 antibody, anti-CD28 antibody, anti-CD3 antibody, anti-PD-1 antibody, anti-PD-L1 antibody), antibodies that enhance immune cell function (e.g., anti-GITR antibody or anti- OX-40 antibody), toll-like receptors (e.g. TLR4, TLR7, TLR9), soluble ligands (e.g. GITRL or OX-40L), or members of the B7 family (e.g. CD80, CD86). is not limited to An additional immunotherapeutic agent (eg, an immune response stimulating agent) can be administered before the TIGIT binding agent is administered, at the same time as the TIGIT binding agent is administered, and/or after the TIGIT binding agent is administered. can. Also provided are pharmaceutical compositions comprising a TIGIT binding agent and an additional immunotherapeutic agent (eg, an immune response stimulating agent). In some embodiments, an immunotherapeutic agent comprises one, two, three, or more immunotherapeutic agents. In some embodiments, the immune response stimulating agent comprises one, two, three, or more immune response stimulating agents.

In some aspects of the methods described herein, the additional therapeutic agent is an antibody that is an immune checkpoint inhibitor. In some embodiments, the immune checkpoint inhibitor is an anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA4 antibody, anti-CD28 antibody, anti-LAG3 antibody, anti-TIM3 antibody, anti-GITR antibody, or anti-OX-40 is an antibody. In some embodiments, the immune checkpoint inhibitor is an anti-4-1BB antibody. In some embodiments, the additional therapeutic agent is an anti-PD-1 antibody selected from the group consisting of nivolumab (Opdivo), pembrolizumab (Keytruda), or pidilzumab. In some embodiments, the additional therapeutic agent is an anti-PD-1 antibody selected from the group consisting of MEDI0680, REGN2810, BGB-A317, and PDR001. In some embodiments, the additional therapeutic agent is an anti-PD-L1 antibody selected from the group consisting of BMS935559 (MDX-1105), atezolizumab (Tecentriq), durvalumab (MEDI4736), or avelumab (MSB0010718C). In some embodiments, the additional therapeutic agent is an anti-CTLA-4 antibody selected from the group consisting of ipilimumab (Yervoy) or tremelimumab. In some embodiments, the additional therapeutic agent is an anti-LAG-3 antibody selected from the group consisting of BMS-986016 and LAG525. In some embodiments, the additional therapeutic agent is an anti-OX-40 antibody selected from the group consisting of MEDI6469, MEDI0562, and MOXR0916. In some embodiments, the additional therapeutic agent is an anti-4-1BB antibody selected from the group consisting of PF-05082566.

Additionally, treatment with a TIGIT binding agent includes concomitant treatment with other biomolecules, such as one or more cytokines (eg, lymphokines, interleukins, interferons, tumor necrosis factors, and/or growth factors). may be accompanied by surgical removal of the tumor, removal of cancer cells, or any other therapy deemed necessary by the treating physician.

In some embodiments, the TIGIT binding agent is adrenomedullin (AM), angiopoietin (Ang), BMP, BDNF, EGF, erythropoietin (EPO), FGF, GDNF, G-CSF, GM-CSF, GDF9, HGF, HDGF, IGF, migration stimulator, myostatin (GDF-8), NGF, neurotrophin, PDGF, thrombopoietin, TGF-α, TGF-β, TNF-α, VEGF, PlGF, γ-IFN, IL-1, IL-2 , IL-3, IL-4, IL-5, IL-6, IL-7, IL-12, IL-15, and IL-18. . In some embodiments, the TIGIT binding agent can be administered in combination with a biomolecule selected from the group consisting of macrophage colony stimulating factor (M-CSF) and stem cell factor (SCF).

In some embodiments of the methods described herein, treatment with a TIGIT binding agent includes surgical removal of tumors, removal of cancer cells, or other treatment deemed necessary by the treating physician. May be accompanied by any surgical therapy.

In certain embodiments of the methods described herein, treatment involves concomitant administration of a TIGIT binding agent and radiation therapy. Treatment with a TIGIT binding agent may occur prior to administration of radiation therapy, concurrently with administration of radiation therapy, or after administration of radiation therapy. Dosage regimens for such radiotherapy can be determined by one skilled in the art.

Co-administration may involve co-administration in a single pharmaceutical formulation, or co-administration using separate formulations, in either order, but generally all of the active agents have their biological activity may include sequential administration within a period of time such that the

It is understood that the combination of the TIGIT binding agent and the at least one additional therapeutic agent may be administered in any order or may be administered simultaneously. In some embodiments, the TIGIT binding agent is administered to a patient who has been previously treated with a second/different therapeutic agent. In certain other embodiments, the TIGIT binding agent and the second additional therapeutic agent are administered substantially simultaneously or simultaneously. For example, a subject may be given a TIGIT binding agent while undergoing a course of treatment with an additional therapeutic agent (eg, chemotherapy). In certain embodiments, the TIGIT binding agent will be administered within one year of treatment with the second therapeutic agent. In certain other embodiments, the TIGIT binding agent will be administered within 10 months, 8 months, 6 months, 4 months, or 2 months of treatment with the second therapeutic agent. In certain other embodiments, the TIGIT-binding agent will be administered within 4 weeks, 3 weeks, 2 weeks, or 1 week of any treatment with a second anti-cancer agent. In some embodiments, the agent is administered within 5 days, 4 days, 3 days, 2 days, or 1 day of any treatment with the second anticancer agent. Further, it is understood that the two (or more) agents or treatments may be administered to the subject within hours or minutes (ie, substantially simultaneously).

When treating disease, the appropriate dosage of a TIGIT binding agent depends on the type of disease to be treated, the severity and course of the disease, the responsiveness of the disease, whether the agent is administered therapeutically or prophylactically. Whether or not depends on previous therapy, patient history, etc. All of this is at the discretion of the treating physician. The TIGIT binding agent may be administered at once, or may be administered over a series of treatments lasting from several days to several months, and administered until cured or until the disease state is reduced (eg, tumor size is reduced). may In some embodiments, the TIGIT binding agent is administered over a course of treatments until disease progression. In some embodiments, optimal dosing regimens are calculated from measurements of drug accumulation in the patient's body and vary according to the relative potency of individual agents. The administering physician can determine optimum dosages, dosing methodologies and repetition rates.

In certain embodiments of the methods described herein, the dosage of the TIGIT binding agent is 0.01 μg to 100 mg/kg body weight, 0.1 μg to 100 mg/kg body weight, 1 μg to 100 mg/kg body weight, 1 mg to 100 mg/kg Body weight, 1 mg to 80 mg/kg body weight, 10 mg to 100 mg/kg body weight, 10 mg to 75 mg/kg body weight, or 10 mg to 50 mg/kg body weight. In certain embodiments, the TIGIT binding agent is about 0.1 mg/kg, about 0.3 mg/kg, about 1 mg/kg, about 1.5 mg/kg, about 2 mg/kg, about 2.5 mg/kg, about 3 mg/kg, A dose of about 5 mg/kg, about 7.5 kg/kg, about 10 mg/kg, about 12 mg/kg, about 12.5 mg/kg, about 15 mg/kg, about 17.5 mg/kg, about 20 mg/kg, or about 25 mg/kg administered to the subject at In certain embodiments, the dose of TIGIT binding agent is from about 0.1 mg to about 20 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 0.3 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 1 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 1.5 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 2 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 2.5 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 3 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 5 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 7.5 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 10 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 12.5 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 15 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 17.5 mg/kg. In some embodiments, the dose of TIGIT binding agent is about 20 mg/kg. In certain embodiments, doses can be given once or more daily, weekly, monthly, or yearly. In certain embodiments, the TIGIT-binding agent is given once a week, once every two weeks, once every three weeks, or once every four weeks.

In some embodiments, the TIGIT binding agent may be administered in a large initial "loading" dose, followed by one or more smaller doses. In some embodiments, dosing frequency may also vary. In some embodiments, the dosing regimen is to administer an initial dose, followed by a further dose (or " administering a "maintenance" dose). For example, a dosing regimen may include administering an initial loading dose followed by weekly maintenance doses, eg, half the initial dose. Alternatively, the dosing regimen may comprise administering an initial loading dose followed by administration of a maintenance dose, eg, half of the initial dose every other week. Alternatively, the dosing regimen may comprise administering three initial doses over three weeks, followed by a maintenance dose, eg, the same dose every other week.

As is known to those of skill in the art, administration of any therapeutic agent can result in side effects and/or toxicity. In some cases, the side effects and/or toxicity are severe enough to prevent administration of a therapeutically effective dose of a particular agent. In some cases, drug therapy must be discontinued and other agents may be tried. However, many agents in the same therapeutic class often exhibit similar side effects and/or toxicities. This means that the patient must stop therapy or, if possible, suffer unpleasant side effects associated with the therapeutic agent.

In some embodiments, the dosing regimen may be limited to a certain number of doses or "cycles." In some embodiments, the agent is administered for 3, 4, 5, 6, 7, 8, or more cycles. For example, the agent is administered every 2 weeks for 6 cycles, the agent is administered every 3 weeks for 6 cycles, the agent is administered every 2 weeks for 4 cycles, The agent is administered every 3 weeks for 4 cycles, and so on. Dosage regimens can be determined and subsequently varied by those skilled in the art.

In embodiments, the TIGIT binding agent is administered about once a week, about once every two weeks, about once every three weeks, about once every four weeks, about once every five weeks, about once every six weeks. about once every 8 weeks, about once every 12 weeks, about once every month, about once every 2 months, about once every 3 months, or once longer be done. In certain embodiments, the TIGIT binding agent is about 0.1 mg/kg, about 0.3 mg/kg, about 1 mg/kg, about 1.5 mg/kg, about 2 mg/kg, about 2.5 mg/kg, about 3 mg/kg, A dose of about 5 mg/kg, about 7.5 kg/kg, about 10 mg/kg, about 12 mg/kg, about 12.5 mg/kg, about 15 mg/kg, about 17.5 mg/kg, about 20 mg/kg, or about 25 mg/kg and administered to the subject approximately once every two weeks.

In some embodiments, the TIGIT binding agent is administered to the subject once every two weeks at a dose of about 0.3 mg/kg.

In some embodiments, the TIGIT binding agent is administered to the subject at a dose of about 1 mg/kg once every two weeks.

In some embodiments, the TIGIT binding agent is administered to the subject once every two weeks at a dose of about 3 mg/kg.

In some embodiments, the TIGIT binding agent is administered to the subject once every two weeks at a dose of about 10 mg/kg.

In some embodiments, the TIGIT binding agent is administered to the subject once every two weeks at a dose of about 15 mg/kg.

In certain embodiments, the subject has undergone prior treatment with a PD-1 antagonist and/or a PD-L1 antagonist. In certain embodiments, the subject has a histologically confirmed advanced recurrent or refractory solid tumor.

In certain embodiments, the subject has head and neck cancer, esophageal/gastroesophageal cancer, gastric cancer, colorectal cancer, anal cancer, hepatocellular/liver cancer, cervical cancer, lung cancer, Melanoma, Merkel cell carcinoma, renal cell/kidney cancer, bladder cancer, ovarian cancer, pancreatic cancer, endometrial cancer, anal cancer, triple-negative breast cancer, known MSI solid tumors ( (including MSI CRC), or have MSS colorectal cancer.

The present invention provides methods of administering a TIGIT binding agent to a subject comprising using an intermittent dosing strategy to administer one or more agents. Employing an intermittent dosing strategy may reduce side effects and/or toxicity associated with the administration of agents, chemotherapeutic agents, and the like. In some embodiments, a method for inhibiting tumor growth and/or treating cancer in a human subject comprises combining a therapeutically effective dose of a TIGIT binding agent with a therapeutically effective dose of a chemotherapeutic agent. Administration to a subject in combination, wherein one or both agents are administered according to an intermittent dosing strategy. In some embodiments, the intermittent dosing strategy comprises administering an initial dose of the TIGIT binding agent to the subject and administering subsequent doses of the TIGIT binding agent approximately once every two weeks. In some embodiments, the intermittent dosing strategy comprises administering an initial dose of the TIGIT binding agent to the subject and administering subsequent doses of the TIGIT binding agent approximately once every three weeks. In some embodiments, the intermittent dosing strategy comprises administering an initial dose of the TIGIT binding agent to the subject and administering subsequent doses of the TIGIT binding agent approximately once every four weeks. In some embodiments, the TIGIT binding agent is administered using an intermittent dosing strategy and the second agent is administered weekly.

The present invention provides methods of using compositions comprising TIGIT binding agents. The invention also provides methods described herein using pharmaceutical compositions comprising a TIGIT binding agent and a pharmaceutically acceptable vehicle. In some embodiments, the pharmaceutical composition is useful in immunotherapy. In some embodiments, the compositions are useful for inhibiting tumor growth. In some embodiments, the pharmaceutical composition is useful for inhibiting tumor growth in a subject (eg, human patient). In some embodiments, the compositions are useful for treating cancer. In some embodiments, the pharmaceutical composition is useful for treating cancer in a subject (eg, a human patient).

By combining a purified TIGIT binding agent and a pharmaceutically acceptable vehicle (eg, carrier or excipient), formulations for storage and use are prepared. Those skilled in the art generally consider pharmaceutically acceptable carriers, excipients, and/or stabilizers to be inactive ingredients of a formulation or pharmaceutical composition.

Suitable pharmaceutically acceptable vehicles include non-toxic buffers such as phosphate, citric acid and other organic acids; salts such as sodium chloride; antioxidants including ascorbic acid and methionine; preservatives. , e.g. octadecyldimethylbenzylammonium chloride, hexamethonium chloride, benzalkonium chloride, benzethonium chloride, phenol, butyl or benzyl alcohol, alkylparabens, e.g. methylparaben or propylparaben, catechol, resorcinol, cyclohexanol, 3-pen low molecular weight polypeptides (eg, less than about 10 amino acid residues); proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers, such as polyvinylpyrrolidone; amino acids, such as glycine; , glutamine, asparagine, histidine, arginine, or lysine; carbohydrates, such as monosaccharides, disaccharides, glucose, mannose, or dextrins; chelating agents, such as EDTA; sugars, such as sucrose, mannitol, trehalose, or sorbitol; such as sodium; metal complexes such as Zn-protein complexes; and nonionic surfactants such as TWEEN or polyethylene glycol (PEG). (Remington: The Science and Practice of Pharmacy, 22nd ^Edition , 2012, Pharmaceutical Press, London.).

The pharmaceutical compositions of this invention can be administered in a number of ways for local or systemic treatment. In some embodiments, the administration comprises (i) topical administration by epidermal or transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids, and powders; (ii) administration by nebulizer. pulmonary, intratracheal, and intranasal administration by inhalation or insufflation of powders or aerosols; (iii) oral administration; parenteral administration, including intramuscular administration (eg, injection or infusion); or intracranial administration (eg, intrathecal or intracerebroventricular administration).

A therapeutic formulation may be in unit dosage form. Such formulations include tablets, pills, capsules, powders, granules, solutions or suspensions in water or non-aqueous media, or suppositories. In solid compositions such as tablets, the principal active ingredient is mixed with a pharmaceutical carrier. Traditional tableting ingredients include cornstarch, lactose, sucrose, sorbitol, talc, stearic acid, magnesium stearate, dicalcium phosphate, or gums, and diluents such as water. These can be used to form solid preformulation compositions containing intimate mixtures of a compound of the present invention, or non-toxic pharmaceutically acceptable salts thereof. The solid preformulation composition is then subdivided into unit dosage forms of the type described above. Tablets, pills, etc. of the formulation or composition may be coated or otherwise compounded to provide a dosage form that confers the advantage of prolonged action. For example, a tablet or pill may have an inner composition coated by an outer component. Additionally, the two components may be separated by an enteric layer that may act to resist disintegration, allowing the inner component to pass intact through the stomach or delaying its release. Various materials can be used for such enteric layers or coatings. Such materials include many polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol, and cellulose acetate.

The TIGIT binding agent can also be encapsulated in microcapsules. Such microcapsules are prepared, for example, by coacervation techniques or interfacial polymerization methods, respectively, as described, for example, in Remington, The Science and Practice of Pharmacy, 22nd ^Edition , 2012, Pharmaceutical Press, London. , hydroxymethylcellulose or gelatin-microcapsules and poly(methyl methacrylate) microcapsules in colloidal drug delivery systems (e.g., liposomes, albumin microspheres, microemulsions, nanoparticles, and nanocapsules) or in macroemulsions. insert.

In certain embodiments, the pharmaceutical formulation includes a TIGIT binding agent complexed with liposomes. Methods of producing liposomes are known to those skilled in the art. For example, some liposomes can be made by reverse-phase evaporation using a lipid composition containing phosphatidylcholine, cholesterol, and PEG-derivatized phosphatidylethanolamine (PEG-PE). Liposomes with the desired diameter can be obtained by extruding liposomes through filters of defined pore size.

In certain embodiments, sustained release preparations are produced that include the TIGIT binding agent. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the agent, which matrices are in the form of shaped articles (eg films or microcapsules). ing. Examples of sustained release matrices include polyesters, hydrogels such as poly(2-hydroxyethyl-methacrylate) or poly(vinyl alcohol), polylactic acid, copolymers of L-glutamic acid and 7-ethyl-L-glutamate, non-degradable Ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as LUPRON DEPOT™ (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), sucrose acetate isobutyrate, and poly-D Contains -(-)-3-hydroxybutyric acid.

III.TIGIT binders
T-cell immunoreceptors with Ig and ITIM domains (TIGIT) are type I transmembrane glycoproteins containing immunoglobulin variable (IgV) domains. TIGIT belongs to the poliovirus receptor (PVR) family and binds with high affinity to the poliovirus receptor (PVR; CD155) and with lower affinity to PVRL-2 (CD112) and PVRL-3 (CD113). Join. TIGIT is expressed on T cells, including regulatory T cells (Treg) and memory T cells, as well as on NK cells, and is upregulated after naive CD4+ T cells are activated. The full-length amino acid (aa) sequence of human TIGIT (UniProtKB No.Q495A1) is known in the art and is provided herein as SEQ ID NO:1. As used herein, references to amino acid positions refer to the numbering of the full length amino acid sequence including the signal sequence.

The invention provides methods of using agents that specifically bind to TIGIT. These agents are referred to herein as "TIGIT binders." In some embodiments, the TIGIT binding agent is an antibody. In some embodiments, the TIGIT binding agent is a polypeptide. In certain embodiments, the TIGIT binding agent binds to mouse TIGIT. In certain embodiments, the TIGIT binding agent binds to human TIGIT. In certain embodiments, the TIGIT binding agent binds to mouse TIGIT and human TIGIT. In some embodiments, the TIGIT binding agent binds to human TIGIT and not to mouse TIGIT. Non-limiting examples of TIGIT binding agents are described, for example, in International Patent Application No. PCT/US2016/034549; International Patent Publication Nos. WO2016/106302 and WO2016/028656; .

In some embodiments, the agent binds to TIGIT and interferes with the interaction of TIGIT with a second protein. In some embodiments, the agent binds to TIGIT and interferes with the interaction of TIGIT and PVR. In some embodiments, the agent binds to TIGIT and interferes with the interaction between TIGIT and PVRL2. In some embodiments, the agent binds to TIGIT and interferes with the interaction between TIGIT and PVRL3. In some embodiments, the agent binds to TIGIT and the agent disrupts the binding of TIGIT to PVR and/or disrupts PVR activation of TIGIT signaling.

In certain embodiments, the TIGIT binding agent is an antibody or fragment thereof that specifically binds to the extracellular domain of TIGIT. In certain embodiments, the TIGIT binding agent is an antibody or fragment thereof that specifically binds to the extracellular domain of mouse TIGIT. In certain embodiments, the TIGIT binding agent is an antibody or fragment thereof that specifically binds to the extracellular domain of human TIGIT. In certain embodiments, the TIGIT binding agent is an antibody or fragment thereof that specifically binds to the extracellular domain of mouse TIGIT and human TIGIT. In some embodiments, the TIGIT binding agent is an antibody that specifically binds to the Ig-like domain of TIGIT. In some embodiments, the TIGIT binding agent is an antibody that specifically binds to the IgV domain of TIGIT. In some embodiments, the TIGIT binding agent is an antibody that binds within amino acids 22-141 of human TIGIT. In some embodiments, the TIGIT binding agent is an antibody that binds within amino acids 22-141 of SEQ ID NO:1. In some embodiments, the agent binds within amino acids 22-124 of human TIGIT. In some embodiments, the agent binds within amino acids 22-124 of SEQ ID NO:1. In certain embodiments, the TIGIT binding agent binds within SEQ ID NO:3 or a fragment thereof. In some embodiments, the TIGIT binding agent is an antibody that binds within amino acids 50-124 of human TIGIT. In some embodiments, the TIGIT binding agent is an antibody that binds within amino acids 50-124 of SEQ ID NO:1. In certain embodiments, the TIGIT binding agent binds within SEQ ID NO:3 or a fragment thereof.

In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope comprising amino acids in SEQ ID NO:27. In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope comprising amino acids in SEQ ID NO:28. In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope comprising amino acids in SEQ ID NO:27 and SEQ ID NO:28. In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope comprising amino acids Q62 and I109 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope comprising amino acids Q62 and T119 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope comprising amino acids Q64 and I109 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope comprising amino acids Q64 and T119 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope comprising amino acids Q62, Q64, and I109 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope comprising amino acids Q62, Q64, and T119 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope comprising amino acids Q62, I109, and T119 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope comprising amino acids Q64, I109, and T119 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope comprising amino acids Q62, Q64, I109, and T119 of SEQ ID NO:1. In some embodiments, the TIGIT binding agent is at least one selected from the group consisting of N58, E60, Q62, Q64, L65, F107, I109, H111, T117, T119, G120, and R121 of SEQ ID NO:1. It is an antibody that binds to an epitope containing three amino acids. In some embodiments, the epitope is a conformational epitope. In some embodiments, the TIGIT binding agent is an antibody that binds to an epitope of SEQ ID NO:1 that does not include amino acid V100.

In certain embodiments, the TIGIT binding agent (e.g., antibody) is about 1 μM or less, about 100 nM or less, about 40 nM or less, about 20 nM or less, about 10 nM or less, about 1 nM or less. Binds TIGIT with a dissociation constant (K _D ) of less than, about 0.1 nM or less, 50 pM or less, 10 pM or less, or 1 pM or less. In some embodiments, the TIGIT binding agent binds TIGIT with a K _D of about 20 nM or less. In some embodiments, the TIGIT binding agent binds TIGIT with a K _D of about 10 nM or less. In some embodiments, the TIGIT binding agent binds TIGIT with a K _D of about 1 nM or less. In some embodiments, the TIGIT binding agent binds TIGIT with a K _D of about 0.5 nM or less. In some embodiments, the TIGIT binding agent binds TIGIT with a K _D of about 0.1 nM or less. In some embodiments, the TIGIT binding agent binds TIGIT with a K _D of about 50 pM or less. In some embodiments, the TIGIT binding agent binds TIGIT with a K _D of about 25 pM or less. In some embodiments, the TIGIT binding agent binds TIGIT with a K _D of about 10 pM or less. In some embodiments, the TIGIT binding agent binds TIGIT with a K _D of about 1 pM or less. In some embodiments, the TIGIT binding agent binds both human and mouse TIGIT with a K _D of about 10 nM or less. In some embodiments, the TIGIT binding agent binds both human and mouse TIGIT with a K _D of about 1 nM or less. In some embodiments, the TIGIT binding agent binds both human and mouse TIGIT with a K _D of about 0.1 nM or less. In some embodiments, the dissociation constant of a binding agent (e.g., antibody) to TIGIT is determined using a TIGIT fusion protein comprising at least a portion of the extracellular domain of the TIGIT protein immobilized on a Biacore chip. is a constant. In some embodiments, the dissociation constant of the binder (e.g., antibody) to TIGIT is the dissociation constant determined using soluble TIGIT protein with the binder captured by an anti-human IgG antibody on a Biacore chip.

In some embodiments, the TIGIT binding agent comprises a first antigen binding site that specifically binds TIGIT and a second antigen binding site that specifically binds a second target. In some embodiments, the TIGIT binding agent is bispecific, comprising a first antigen binding site that specifically binds TIGIT and a second antigen binding site that specifically binds a second target. It is matter. In some embodiments, the TIGIT binding agent binds both TIGIT and the second target with a K _D of about 100 nM or less. In some embodiments, the TIGIT binding agent binds both TIGIT and the second target with a K _D of about 50 nM or less. In some embodiments, the TIGIT binding agent binds both TIGIT and the second target with a K _D of about 20 nM or less. In some embodiments, the TIGIT binding agent binds both TIGIT and the second target with a K _D of about 10 nM or less. In some embodiments, the TIGIT binding agent binds both TIGIT and the second target with a K _D of about 1 nM or less. In some embodiments, the affinity of one antigen binding site may be weaker than the affinity of the other antigen binding site. For example, one antigen binding site may have a K _D of about 1 nM and a second antigen binding site may have a K _D of about 10 nM. In some embodiments, the difference in affinity between the two antigen binding sites is about 2-fold or more, about 3-fold or more, about 5-fold or more, about 8-fold or more, about 10-fold or more. It may be about 15 times or more, about 20 times or more, about 30 times or more, about 50 times or more, or about 100 times or more. Modulation of the affinity of the two antigen-binding sites can affect the biological activity of bispecific antibodies. For example, decreasing the affinity of the antigen binding site for TIGIT or a second target may have desirable effects, such as decreasing toxicity and/or increasing the therapeutic index of the binding agent.

In certain embodiments, the TIGIT binding agent (e.g., antibody) is about 1 μM or less, about 100 nM or less, about 40 nM or less, about 20 nM or less, about 10 nM or less, about 1 nM or less. Binds TIGIT at a ₅₀ % effective concentration (EC50) of less than, or about 0.1 nM or less. In certain embodiments, the TIGIT binding agent is about 1 μM or less, about 100 nM or less, about 40 nM or less, about 20 nM or less, about 10 nM or less, about 1 nM or less, or about 0.1 Binds human TIGIT at 50% effective concentration (EC ₅₀ ) of nM or less. In certain embodiments, the TIGIT binding agent binds mouse TIGIT and/or mouse _TIGIT with an EC50 of about 40 nM or less, about 20 nM or less, about 10 nM or less, about 1 nM or less, or about 0.1 nM or less. or bind to human TIGIT.

In certain embodiments, the TIGIT binding agent is an antibody. In some embodiments, said antibody is a recombinant antibody. In some embodiments, said antibody is a monoclonal antibody. In some embodiments, said antibody is a chimeric antibody. In some embodiments, said antibody is a humanized antibody. In some embodiments, said antibody is a human antibody. In some embodiments, the antibody is an IgA, IgD, IgE, IgG, or IgM antibody. In certain embodiments, said antibody is an IgG1 antibody. In certain embodiments, said antibody is an IgG2 antibody. In some embodiments, said antibody is an IgG4 antibody. In certain embodiments, said antibody is an antibody fragment that comprises an antigen-binding site. In some embodiments, said antibody is a bispecific or multispecific antibody. In some embodiments, said antibody is a monovalent antibody. In some embodiments, said antibody is a monospecific antibody. In some embodiments, said antibody is a bivalent antibody. In some embodiments, the antibody is conjugated to a cytotoxic moiety. In some embodiments, the antibody is isolated. In some embodiments, the antibody is substantially pure.

In some embodiments, the TIGIT binding agent is a polyclonal antibody. Polyclonal antibodies can be prepared by any known method. In some embodiments, polyclonal antibodies are administered to animals (eg, rabbits, rats, mice, goats, donkeys) using multiple subcutaneous or intraperitoneal injections with an antigen of interest (eg, a purified peptide fragment). , full-length recombinant protein, or fusion protein). The antigen can optionally be conjugated to a carrier such as keyhole limpet hemocyanin (KLH) or serum albumin. Antigen (with or without carrier protein) is diluted in sterile saline and usually combined with an adjuvant (eg, complete Freund's adjuvant or incomplete Freund's adjuvant) to form a stable emulsion. After sufficient time has elapsed, polyclonal antibodies are recovered from the immunized animal, usually from the blood or ascites fluid. Polyclonal antibodies can be purified from serum or ascites fluid according to standard methods in the art, including, but not limited to, affinity chromatography, ion exchange chromatography, gel electrophoresis, and dialysis.

In some embodiments, the TIGIT binding agent is a monoclonal antibody. Monoclonal antibodies can be prepared using hybridoma methods known to those of skill in the art. In some embodiments using hybridoma technology, mice, rats, rabbits, hamsters, or other suitable host animals are immunized as described above to elicit the production of antibodies that specifically bind the immunizing antigen. be done. In some embodiments, lymphocytes can be immunized in vitro. In some embodiments, the immunizing antigen can be a human protein or fragment thereof. In some embodiments, the immunizing antigen can be a mouse protein or fragment thereof.

After immunization, lymphocytes are isolated and fused with a suitable myeloma cell line using, for example, polyethylene glycol. Hybridoma cells are selected using specialized media as known in the art, and unfused lymphocytes and myeloma cells do not survive the selection process. Hybridomas producing monoclonal antibodies directed specifically against a selected antigen are subject to immunoprecipitation, immunoblotting, and in vitro binding assays (e.g., flow cytometry, FACS, ELISA, and radioimmunoassay), but It can be identified by various methods, including but not limited to these. Hybridomas may be grown in vitro culture using standard methods, or in vivo as ascites tumors in animals. Monoclonal antibodies can be purified from culture medium or ascites fluid according to methods standard in the art, including, but not limited to, affinity chromatography, ion exchange chromatography, gel electrophoresis, and dialysis.

In certain embodiments, monoclonal antibodies can be made using recombinant DNA methods, as known to those of skill in the art. Polynucleotides encoding monoclonal antibodies are isolated from mature B cells or hybridoma cells, for example, by RT-PCR using oligonucleotide primers that specifically amplify the genes encoding the antibody heavy and light chains. , the sequences of which are determined using standard techniques. The isolated polynucleotides encoding the heavy and light chains are then cloned into a suitable expression vector, which is transfected into a host cell, such as E. coli, which otherwise does not produce immunoglobulin proteins. (E. coli), monkey COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells produce monoclonal antibodies.

In certain other embodiments, recombinant monoclonal antibodies or fragments thereof can be isolated from phage display libraries expressing variable domains or CDRs of the desired species.

Polynucleotides encoding monoclonal antibodies can be modified to produce alternative antibodies, for example, by using recombinant DNA techniques. In some embodiments, the constant domains of the light and heavy chains, e.g., of a murine monoclonal antibody, are combined with constant regions, e.g., the constant regions of a human antibody, or Non-immunoglobulin polypeptides can be substituted to create fusion antibodies. In some aspects, the constant regions are truncated or removed to generate the desired antibody fragment of the monoclonal antibody. Site-directed or high-density mutagenesis of the variable regions can be used to optimize specificity, affinity, etc. of monoclonal antibodies.

In some embodiments, the TIGIT binding agent is a humanized antibody. Typically, humanized antibodies are isolated from non-human species (e.g., mouse, rat) in which the amino acid residues of the CDRs have the desired specificity, affinity, and/or binding capacity, using methods known to those of skill in the art. , rabbit, hamster, etc.) are replaced by amino acid residues from the CDRs. In some embodiments, some of the framework variable region amino acid residues of the human immunoglobulin are replaced with corresponding amino acid residues in an antibody derived from a non-human species. In some embodiments, the humanized antibody is further modified by substitution of additional residues in the framework variable region and/or within the replaced non-human residues to improve antibody specificity, affinity, and/or Capabilities can be further improved and optimized. Generally, a humanized antibody comprises a variable domain region comprising all or substantially all CDRs that correspond to those of a non-human immunoglobulin, whereas all or substantially all framework regions correspond to those of a human immunoglobulin. It is the framework region of the sequence. In some embodiments, the framework regions are those of a human consensus immunoglobulin sequence. In some embodiments, the humanized antibody also will comprise at least a portion of an immunoglobulin constant region or domain (Fc), typically that of a human immunoglobulin. can. In certain embodiments, such humanized antibodies are used therapeutically as they may reduce antigenicity and HAMA (human anti-mouse antibody) responses when administered to a human subject.

In certain embodiments, the TIGIT binding agent is a human antibody. Human antibodies can be prepared directly using various techniques known in the art. In some embodiments, human antibodies may be generated from immortalized human B lymphocytes immunized in vitro or from lymphocytes isolated from the immunized individual. In either case, cells can be generated and isolated that produce antibodies directed against the target antigen. In some embodiments, human antibodies can be selected from phage libraries that express human antibodies. Alternatively, phage display technology can be used to generate human antibodies and antibody fragments in vitro from immunoglobulin variable domain gene repertoires derived from unimmunized donors. Techniques for making and using antibody phage libraries are well known in the art. Once antibodies are identified, affinity maturation strategies known in the art, including but not limited to chain shuffling and site-directed mutagenesis, may be used to generate high affinity human antibodies.

In some embodiments, human antibodies can be produced in transgenic mice that contain human immunoglobulin loci. When these mice are immunized, they are capable of producing a full repertoire of human antibodies in the absence of endogenous immunoglobulin production.

In some embodiments, the TIGIT binding agent is a bispecific antibody. Accordingly, the present invention encompasses bispecific antibodies that specifically recognize TIGIT and at least one additional target. Bispecific antibodies are capable of specifically recognizing and binding at least two different antigens or epitopes. Different epitopes may be present within the same molecule (eg two epitopes in TIGIT) or in different molecules (eg one epitope in TIGIT and one epitope in different proteins). In some embodiments, bispecific antibodies are more effective than individual antibodies or as a combination of multiple antibodies. In some embodiments, bispecific antibodies are less toxic compared to individual antibodies or compared to combinations of multiple antibodies. It is known to those skilled in the art that any therapeutic agent can have a unique pharmacokinetic (PK) (eg, circulating half-life). In some embodiments, bispecific antibodies are capable of co-generating the PKs of two active binding agents. In this case, the PK profiles of the two individual binding agents are different. In some embodiments, bispecific antibodies are capable of concentrating the actions of two agents into a common region (eg, a tumor and/or tumor microenvironment). In some embodiments, bispecific antibodies are capable of concentrating the actions of two agents on a common target (eg, a tumor or tumor cell). In some embodiments, bispecific antibodies are capable of targeting the actions of two agents to multiple biological pathways or functions. In some embodiments, bispecific antibodies have the ability to target and bring two different cells close together (eg, immune cells and tumor cells).

In some embodiments, bispecific antibodies are monoclonal antibodies. In some embodiments, bispecific antibodies are humanized antibodies. In some embodiments, the bispecific antibody is a human antibody. In some embodiments, the bispecific antibody is an IgG1 antibody. In some embodiments, the bispecific antibody is an IgG2 antibody. In some embodiments, the bispecific antibody is an IgG4 antibody. In some embodiments, bispecific antibodies have reduced toxicity and/or side effects. In some embodiments, bispecific antibodies have reduced toxicity and/or side effects compared to a mixture of two individual antibodies or the antibody as a single agent. In some embodiments, bispecific antibodies have large therapeutic indices. In some embodiments, bispecific antibodies have increased therapeutic indices compared to a mixture of two individual antibodies or the antibody as a single agent.

In some embodiments, the antibody is a TIGIT as well as a second antigenic target, such as an effector molecule (eg, CD2, CD3, CD28, CTLA4, PD-1, PD-L1, CD80, or CD86) on an immune cell. or can specifically recognize and bind Fc receptors (eg, CD64, CD32, or CD16). In some embodiments, the antibodies can be used to direct cytotoxic agents to cells expressing a particular target antigen. These antibodies possess an antigen-binding arm and an arm that binds a cytotoxic or radionuclide chelating agent, eg, EOTUBE, DPTA, DOTA, or TETA.

Techniques for making bispecific antibodies are known to those of skill in the art. In some embodiments, bispecific antibodies comprise heavy chain constant regions with altered amino acids that are part of the interface between the two heavy chains. In some embodiments, bispecific antibodies can be generated using a "knob-into-hole" strategy. In some cases, the terms "knob" and "hole" are replaced with the terms "protrusion" and "cavity." In some embodiments, bispecific antibodies may contain variant hinge regions that are incapable of forming disulfide bonds between heavy chains. In some embodiments, modifications may include amino acid changes that alter electrostatic interactions. In some embodiments, modifications may include amino acid changes that alter hydrophobic/hydrophilic interactions.

Bispecific antibodies can be intact antibodies or antibody fragments that contain the antigen-binding site. Antibodies with more than two valencies are also contemplated. For example, trispecific antibodies can be prepared. Thus, in certain embodiments, antibodies to TIGIT are multispecific.

In certain embodiments, the antibodies (or other polypeptides) described herein can be monospecific. In certain embodiments, each of the one or more antigen binding sites comprised by an antibody is capable of binding (or binds to a homologous epitope on TIGIT) a homologous epitope on TIGIT.

In certain embodiments, the TIGIT binding agent is an antibody fragment. Antibody fragments may have different functions or abilities than intact antibodies. For example, antibody fragments can have high tumor penetrance. Various techniques are known for generating antibody fragments, including, but not limited to, proteolytic digestion of intact antibodies. In some embodiments, antibody fragments include F(ab')2 fragments produced by pepsin digestion of the antibody molecule. In some embodiments, antibody fragments include Fab fragments produced by reducing the disulfide bridges of the F(ab')2 fragment. In other embodiments, antibody fragments include Fab fragments produced by treating an antibody molecule with papain and a reducing agent. In certain embodiments, antibody fragments are produced recombinantly. In some embodiments, antibody fragments include Fv or single-chain Fv (scFv) fragments. Fab, Fv, and scFv antibody fragments can be expressed in and secreted from E. coli or other host cells, allowing the production of large amounts of these fragments. . In some embodiments, antibody fragments are isolated from the antibody phage libraries discussed herein. For example, methods for constructing Fab expression libraries can be used to rapidly and effectively identify monoclonal Fab fragments with the desired specificity for TIGIT or its derivatives, fragments, analogs, or homologs. can. In some embodiments, the antibody fragment is a linear antibody fragment. In certain embodiments, antibody fragments are monospecific or bispecific. In certain embodiments, the TIGIT binding agent is a scFv. Various techniques can be used to generate single chain antibodies specific for TIGIT.

In some embodiments, particularly antibody fragments, the antibody is modified to alter (eg, lengthen or shorten) the serum half-life of the antibody. This can be done, for example, by incorporating a salvage receptor binding epitope into the antibody fragment, by mutating the appropriate region in the antibody fragment, or incorporating the epitope into a peptide tag, which is then transformed into ( For example, by DNA synthesis or peptide synthesis), this can be achieved by fusing to the antibody fragment at either end or in the middle.

Heteroconjugate antibodies are also within the scope of the invention. Heteroconjugate antibodies are composed of two covalently joined antibodies. For example, such antibodies have been proposed to target immune cells to unwanted cells. It is also contemplated that heteroconjugate antibodies can be prepared in vitro using known methods in synthetic protein chemistry, including methods involving cross-linking agents. For example, immunotoxins can be constructed using a disulfide exchange reaction or by forming a thioether bond. Examples of suitable reagents for this purpose include iminothiolate and methyl-4-mercaptobutyrimidate.

For the purposes of the present invention, it should be understood that a modified antibody may comprise any type of variable region that associates said antibody with a target (ie, TIGIT). In this regard, the variable region may comprise any type of mammal capable of initiating a humoral response and being induced to generate an immunoglobulin against a desired antigen, including It may be derived from any type of mammal that can be induced to develop immunoglobulins. Thus, the variable regions of the modified antibody can be, for example, variable regions derived from humans, mice, rats, rabbits, non-human primates (eg, cynomolgus monkeys, macaques, etc.), or rabbits. In some embodiments, both the variable and constant regions of the modified immunoglobulin are human. In other embodiments, the variable region of a compatible antibody (usually derived from a non-human source) may be engineered to improve the binding properties of the molecule or to reduce the immunogenicity of the molecule; May be specially tailored. In this regard, variable regions useful in the present invention may be humanized or otherwise altered by the inclusion of imported amino acid sequences.

In certain embodiments, the variable domains in both the heavy and light chains are altered by at least partial exchange of one or more CDRs and, optionally, partial framework region exchanges and sequence modifications. and/or altered by change. The CDRs may be from the same class or even subclass of antibody from which the framework regions were obtained, although the CDRs may be from a different class of antibody, often from a different species. It is envisioned that it may be obtained from an antibody that In order to transfer the antigen-binding capacity of one variable domain to another, it may not be necessary to replace all CDRs with all CDRs from a donor variable region. Conversely, it may be necessary to transfer only those residues necessary to maintain antigen binding site activity.

Despite the changes to the variable region, one skilled in the art will appreciate that the modified antibodies of the present invention exhibit elevated tumor cytotoxicity when compared to antibodies of about the same immunogenicity containing native or unaltered constant regions. Antibodies in which at least a portion of one or more of the constant region domains have been deleted or otherwise altered to obtain desirable biochemical properties such as localization or prolonged serum half-life (e.g. , full-length antibodies or immunoreactive fragments thereof). In some embodiments, the constant region of the modified antibody comprises a human constant region. Modifications to the constant region compatible with this invention comprise additions, deletions or substitutions of one or more amino acids in one or more domains. The modified antibodies disclosed herein have changes or modifications to one or more of the three heavy chain constant domains (CH1, CH2, or CH3) and/or the light chain constant domain (CL). may contain. In some embodiments, one or more domains are partially or completely deleted from the constant region of the modified antibody. In some embodiments, the modified antibodies comprise domain deleted constructs or variants in which the entire CH2 domain has been removed (ΔCH2 constructs). In some embodiments, the removed constant region domain is typically replaced with a short amino acid spacer (eg, 10 amino acid residues) that provides some of the molecular flexibility conferred by the absent constant region. be.

In some embodiments, modified antibodies are engineered to fuse the CH3 domain directly to the hinge region of the antibody. In other embodiments, peptide spacers are inserted between the hinge region and the modified CH2 and/or CH3 domains. For example, a construct may be expressed in which the CH2 domain has been deleted and a 5-20 amino acid spacer is used to join the remaining CH3 domain (modified or unmodified) to the hinge region. The addition of such spacers can keep the regulatory elements of the constant domain accessible in the free state or keep the hinge region flexible. However, it should be noted that amino acid spacers have been found to be immunogenic in some cases and may elicit an undesirable immune response against the construct. Thus, in certain aspects, any spacers added to the construct are relatively non-immunogenic such that the desired biological properties of the modified antibody are maintained.

In some embodiments, the modified antibody may have only partial constant domain deletions, or may have only a few amino acid substitutions, or even single amino acid substitutions. For example, single amino acid mutations in selected regions of the CH2 domain may be sufficient to significantly reduce Fc binding. In some embodiments, single amino acid mutations in selected regions of the CH2 domain may be sufficient to significantly reduce Fc binding and enhance cancer cell localization and/or tissue penetration. be. Similarly, it may be desirable to simply delete part of one or more constant region domains that control the particular effector function to be modulated (eg complement C1q binding). Such partial deletions of the constant region may improve selected properties of the antibody (serum half-life) while leaving other desirable functions associated with this constant region domain intact. Furthermore, as noted above, the constant regions of the disclosed antibodies may be altered by one or more amino acid mutations or substitutions that improve the profile of the resulting construct. In this regard, it may be possible to perturb the activity provided by conserved binding sites (eg, Fc binding) while substantially maintaining the composition and immunogenicity profile of the altered antibody. In certain embodiments, the modified antibody will have one or more mutations in the constant region that enhance desirable properties, such as reduced or increased effector function, or more cytotoxic or carbohydrate attachment sites. of amino acids.

Constant regions are known in the art to mediate several effector functions. For example, the binding of the C1 component of complement to the Fc region of an (antigen-bound) IgG or IgM antibody activates the complement system. Complement activation is important in the opsonization and lysis of cellular pathogens. Complement activation also stimulates inflammatory responses and may be involved in autoimmune hypersensitivity. In addition, the Fc region of antibodies can bind to cells expressing Fc receptors (FcR). There are many Fc receptors specific for different classes of antibodies, including IgG (gamma receptors), IgE (epsilon receptors), IgA (alpha receptors), and IgM (mu receptors). Binding of antibodies to Fc receptors on cell surfaces results in phagocytosis and destruction of antibody-coated particles, clearance of immune complexes, lysis of antibody-coated target cells by killer cells (called antibody-dependent cytotoxicity or ADCC), and inflammation. Many important and diverse biological responses are elicited, including mediator release, placental passage, and regulation of immunoglobulin production.

In certain embodiments, the modified antibody results in altered effector function, which in turn affects the biological profile of the administered antibody. For example, in some embodiments, deletion or inactivation of constant region domains (by point mutations or other means) can reduce Fc receptor binding of circulating modified antibodies. In some embodiments, deletion or inactivation of the constant region domains (by point mutations or other means) reduces Fc receptor binding of the circulating modified antibodies, thereby leading to cancer cell localization. There may be increased localization and/or tumor penetrance. In another embodiment, the constant region modifications increase the serum half-life of said antibody. In another embodiment, the constant region alterations shorten the serum half-life of said antibody. In some embodiments, the constant region is modified to eliminate disulfide bonds or oligosaccharide moieties. Modifications of the constant region according to the present invention may be readily made using well known biochemical or molecular engineering techniques.

In certain embodiments, a TIGIT binding agent that is an antibody lacks one or more effector functions. For example, in some embodiments, the antibody lacks ADCC activity and/or lacks complement dependent cytotoxicity (CDC) activity. In certain embodiments, said antibody does not bind to Fc receptors and/or complement factors. In certain embodiments, said antibody lacks effector functions.

The invention further includes recombinant, monoclonal, chimeric, humanized, and human antibodies described herein, or variants and equivalents that are substantially homologous to the antibodies, or antibody fragments thereof. These variants may contain, for example, conservative substitution mutations, ie the substitution of one or more amino acids by similar amino acids.

In certain embodiments, the antibodies described herein are isolated. In certain embodiments, the antibodies described herein are substantially pure.

TIGIT binding agents (eg, antibodies) of the invention can be assayed for specific binding by any method known in the art. Immunoassays that can be used include Biacore analysis, FACS analysis, immunofluorescence, immunocytochemistry, Western blot analysis, radioimmunoassay, ELISA, "sandwich" immunoassay, immunoprecipitation assay, precipitation reaction, gel diffusion precipitation reaction, immunodiffusion. Competitive and non-competitive assay systems using techniques such as, but not limited to, method assays, agglutination assays, complement fixation assays, immunoradiometric assays, fluorescence immunoassays, and protein A immunoassays. Such assays are routine and well known in the art (see, eg, Ausubel et al., Editors, 1994 present, Current Protocols in Molecular Biology, John Wiley & Sons, Inc., New York, NY). See ).

In a non-limiting example, screening for specific binding between antibodies and human TIGIT may be determined using ELISA. ELISA involves preparing an antigen (eg, TIGIT or a fragment thereof), coating wells of a 96-well microtiter plate with the antigen, detecting a detectable compound, such as an enzymatic substrate (eg, horseradish peroxidase or alkaline phosphatase). adding a test antibody conjugated to the well to the well, incubating for a period of time, and detecting the presence of the antibody bound to the antigen. In some embodiments, the test antibody is not conjugated to a detectable compound, instead a secondary antibody (e.g., an anti-Fc antibody) that recognizes the antibody and is conjugated to a detectable compound. is added to the wells. In some embodiments, instead of coating the wells with the antigen, the test antibody may be coated into the wells and the antigen (e.g., TIGIT) added to the wells before being conjugated to a detectable compound. A secondary antibody is added. One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the signal detected as well as other variations of ELISAs known in the art.

In another non-limiting example, specific binding between an antibody and TIGIT may be determined using FACS. The FACS screening assay involves making a cDNA construct that expresses the antigen as a full-length protein (TIGIT) or a fusion protein (e.g., TIGIT-CD4TM), introducing the construct into cells by transfection, and expressing the antigen on the cell surface. allowing, mixing the test antibody with the transfected cells, and incubating for a period of time. Cells bound by the test antibody can be identified using a secondary antibody (eg, PE-conjugated anti-Fc antibody) conjugated to a detectable compound and a flow cytometer. One of ordinary skill in the art would be knowledgeable as to the parameters that can be modified to optimize the signal detected as well as other variations of FACS that can enhance screening (eg, screening for blocking antibodies).

Competitive binding assays can determine the binding affinity and off-rate of antibody-antigen interaction between an antibody or other binding agent and an antigen (eg, TIGIT). One example of a competitive binding assay involves incubation of a labeled antigen (e.g., ³ H or ¹²⁵ I-TIGIT) or fragment or variant thereof with an antibody of interest in the presence of increasing amounts of unlabeled antigen, followed by labeling. A radioimmunoassay that involves detecting antibody bound to an antigen. The affinity and binding off-rate of the antibody for the antigen can be determined from the data by Scatchard plot analysis. In some embodiments, Biacore kinetic analysis is used to determine binding on- and off-rates of antibodies or agents that bind to an antigen (eg, TIGIT). In some embodiments, Biacore kinetic analysis involves analyzing the binding and dissociation of antibodies from chips with antigens (eg, TIGIT) immobilized on the chip surface. In some embodiments, Biacore kinetic analysis comprises analyzing the binding and dissociation of an antigen (eg, TIGIT) from a chip with antibodies (eg, anti-TIGIT antibodies) immobilized on the chip surface.

In certain embodiments of the methods described herein, the invention provides a TIGIT-binding agent (e.g., an antibody) that specifically binds to human TIGIT, wherein the TIGIT-binding agent comprises the CDRs of antibody OMP-313M32 ( See Table 1). Anti-TIGIT antibody OMP-313M32. In some embodiments, the TIGIT binding agent is one or more of the CDRs of OMP-313M32; two or more of the CDRs of OMP-313M32; three or more of the CDRs of OMP-313M32. 4 or more of the OMP-313M32 CDRs; 5 or more of the OMP-313M32 CDRs; or all 6 of the OMP-313M32 CDRs.

(Table 1)

を含む重鎖CDR2、および

を含む重鎖CDR3を含む。一部の態様において、TIGIT結合物質は、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3をさらに含む。一部の態様において、TIGIT結合物質は、KASQDVSTAVA(SEQ ID NO:7)を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、TIGIT結合物質は、(a)TSDYAWN(SEQ ID NO:4)を含む重鎖CDR1、

を含む重鎖CDR2、および

を含む重鎖CDR3;ならびに(b)

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In certain embodiments of the methods described herein, the invention provides a TIGIT binding agent (e.g., an antibody) that specifically binds human TIGIT, wherein the TIGIT binding agent is TSDYAWN (SEQ ID NO:4 ), including the heavy chain CDR1,

heavy chain CDR2 containing, and

containing a heavy chain CDR3 containing In some embodiments, the TIGIT binding agent is

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the TIGIT binding agent comprises a light chain CDR1 comprising KASQDVSTAVA (SEQ ID NO:7), a light chain CDR2 comprising SASYRYT (SEQ ID NO:8), and QQHYSTP (SEQ ID NO:9). Contains light chain CDR3. In some embodiments, the TIGIT binding agent comprises (a) a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

heavy chain CDR2 containing, and

a heavy chain CDR3 comprising; and (b)

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

を含む重鎖CDR2、または1個、2個、3個、もしくは4個のアミノ酸置換を含むその変種;(c)

を含む重鎖CDR3、または1個、2個、3個、もしくは4個のアミノ酸置換を含むその変種;(d)

を含む軽鎖CDR1、または1個、2個、3個、もしくは4個のアミノ酸置換を含むその変種;(e)SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、または1個、2個、3個、もしくは4個のアミノ酸置換を含むその変種;および(f)QQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3、または1個、2個、3個、もしくは4個のアミノ酸置換を含むその変種
を含む。ある特定の態様において、アミノ酸置換は保存的置換である。一部の態様において、置換はヒト化プロセスの一環としてなされる。一部の態様において、置換は生殖系列ヒト化プロセスの一環としてなされる。一部の態様において、置換は結合最適化プロセスの一環としてなされる。 In certain embodiments of the methods described herein, the invention provides a TIGIT binding agent (e.g., an antibody) that specifically binds to human TIGIT, wherein the TIGIT binding agent comprises (a) TSDYAWN (SEQ ID NO:4), or a variant thereof containing 1, 2, 3, or 4 amino acid substitutions; (b)

or a variant thereof containing 1, 2, 3, or 4 amino acid substitutions; (c)

or a variant thereof containing 1, 2, 3, or 4 amino acid substitutions; (d)

or a variant thereof comprising 1, 2, 3, or 4 amino acid substitutions; (e) a light chain CDR2 comprising SASYRYT (SEQ ID NO:8), or 1, 2 , variants thereof containing 3, or 4 amino acid substitutions; and (f) a light chain CDR3 containing QQHYSTP (SEQ ID NO:9), or 1, 2, 3, or 4 amino acid substitutions. including its variants. In certain embodiments, amino acid substitutions are conservative substitutions. In some embodiments, substitutions are made as part of the humanization process. In some embodiments, substitutions are made as part of the germline humanization process. In some embodiments, substitutions are made as part of a joint optimization process.

In certain embodiments of the methods described herein, the invention provides a TIGIT binding agent (e.g., an antibody) that specifically binds to human TIGIT, wherein the TIGIT binding agent comprises SEQ ID NO:10 and at least A heavy chain variable region having about 80% sequence identity and/or a light chain variable region having at least 80% sequence identity with SEQ ID NO:11. In certain embodiments, the TIGIT binding agent is a heavy chain that has at least about 85%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% sequence identity with SEQ ID NO:10. Contains variable regions. In certain embodiments, the TIGIT binding agent is a light chain having at least about 85%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% sequence identity with SEQ ID NO:11 Contains variable regions. In certain embodiments, the TIGIT binding agent has at least about 95% sequence identity with SEQ ID NO:10 and/or at least about 95% sequence identity with SEQ ID NO:11. It contains the light chain variable region. In certain embodiments, the TIGIT binding agent comprises a heavy chain variable region comprising SEQ ID NO:10 and a light chain variable region comprising SEQ ID NO:11. In certain embodiments, the TIGIT binding agent comprises a heavy chain variable region comprising SEQ ID NO:10 and/or a light chain variable region comprising SEQ ID NO:11. In certain embodiments, the TIGIT binding agent comprises a heavy chain variable region consisting essentially of SEQ ID NO:10 and a light chain variable region consisting essentially of SEQ ID NO:11. In certain embodiments, the TIGIT binding agent comprises a heavy chain variable region consisting of SEQ ID NO:10 and a light chain variable region consisting of SEQ ID NO:11.

In certain embodiments of the methods described herein, the TIGIT binding agent comprises a heavy chain variable region comprising SEQ ID NO:10 and a light chain variable region comprising SEQ ID NO:11. In certain embodiments, the TIGIT binding agent comprises a heavy chain variable region consisting essentially of SEQ ID NO:10 and a light chain variable region consisting essentially of SEQ ID NO:11. In certain embodiments, the TIGIT binding agent comprises a heavy chain variable region consisting of SEQ ID NO:10 and a light chain variable region consisting of SEQ ID NO:11.

In certain embodiments of the methods described herein, the invention provides a TIGIT binding agent (e.g., an antibody) that specifically binds to human TIGIT, wherein the TIGIT binding agent comprises SEQ ID NO:13 and at least A heavy chain with 90% sequence identity and/or a light chain with at least 90% sequence identity to SEQ ID NO:15. In some embodiments, the TIGIT binding agent comprises a heavy chain having at least 95% sequence identity with SEQ ID NO:13 and/or a light chain having at least 95% sequence identity with SEQ ID NO:15. . In some embodiments, the TIGIT binding agent comprises a heavy chain comprising SEQ ID NO:13 and/or a light chain comprising SEQ ID NO:15. In some embodiments, the TIGIT binding agent comprises a heavy chain consisting essentially of SEQ ID NO:13 and a light chain consisting essentially of SEQ ID NO:15. In some embodiments, the TIGIT binding agent comprises a heavy chain consisting of SEQ ID NO:13 and a light chain consisting of SEQ ID NO:15.

In certain embodiments of the methods described herein, the invention provides a TIGIT binding agent (e.g., an antibody) that specifically binds to human TIGIT, wherein the TIGIT binding agent comprises SEQ ID NO:17 and at least A heavy chain with 90% sequence identity and/or a light chain with at least 90% sequence identity to SEQ ID NO:15. In some embodiments, the TIGIT binding agent comprises a heavy chain having at least 95% sequence identity with SEQ ID NO:17 and/or a light chain having at least 95% sequence identity with SEQ ID NO:15. . In some embodiments, the TIGIT binding agent comprises a heavy chain comprising SEQ ID NO:17 and/or a light chain comprising SEQ ID NO:15. In some embodiments, the TIGIT binding agent comprises a heavy chain consisting essentially of SEQ ID NO:17 and a light chain consisting essentially of SEQ ID NO:15. In some embodiments, the TIGIT binding agent comprises a heavy chain consisting of SEQ ID NO:17 and a light chain consisting of SEQ ID NO:15.

In certain embodiments of the methods described herein, the TIGIT binding agent comprises the heavy and light chain variable regions of the OMP-313M32 antibody. In some embodiments, the TIGIT binding agent comprises the variable regions of the OMP-313M32 antibody, wherein the heavy and/or light chain variable regions from the OMP-313M32 antibody have been affinity matured. In certain embodiments, the TIGIT binding agent comprises the heavy and light chains of the OMP-313M32 antibody (with or without leader sequence). In certain embodiments, the TIGIT binding agent is the OMP-313M32 antibody. In certain embodiments, the TIGIT binding agent comprises the heavy chain variable region of the OMP-313M32 antibody as part of an IgG1 heavy chain, IgG2 heavy chain, or IgG4 heavy chain. In certain embodiments, the TIGIT binding agent comprises the heavy chain variable region of the OMP-313M32 antibody as part of a human IgG1 heavy chain. In certain embodiments, the TIGIT binding agent comprises the heavy chain variable region of the OMP-313M32 antibody as part of a human IgG2 heavy chain. In certain embodiments, the TIGIT binding agent comprises the heavy chain variable region of the OMP-313M32 antibody as part of a human IgG4 heavy chain. In certain embodiments, a TIGIT binding agent comprising the heavy chain variable region of the OMP-313M32 antibody as part of a human IgG4 heavy chain is referred to as the OMP-313M33 antibody.

In certain embodiments of the methods described herein, the TIGIT binding agent comprises antibody 313M32, consists essentially of antibody OMP-313M32, or consists of antibody OMP-313M32. In certain embodiments, the TIGIT binding agent comprises a variant of antibody OMP-313M32, consists essentially of a variant of antibody OMP-313M32, or consists of a variant of antibody OMP-313M32. In certain embodiments, the TIGIT binding agent comprises, consists essentially of, or consists of antibody OMP-313M33.

In some embodiments of the methods described herein, the TIGIT-binding agent was obtained from the American Type Culture Collection (ATCC), 10801 University Boulevard, Manassas, VA, USA on Aug. 11, 2015 under the provisions of the Budapest Treaty. It contains the heavy chain variable region encoded by the plasmid deposited at PTA-122346 and designated PTA-122346. In some embodiments, the TIGIT binding agent is encoded by a plasmid designated PTA-122347 deposited at ATCC, 10801 University Boulevard, Manassas, VA, USA on Aug. 11, 2015 under the provisions of the Budapest Treaty. comprising a light chain variable region that is In some embodiments, the TIGIT binding agent is a heavy chain variable region encoded by a plasmid deposited with the ATCC and designated PTA-122346 and a plasmid deposited with the ATCC and designated PTA-122347. and a light chain variable region. In some embodiments, the TIGIT binding agent comprises a heavy chain comprising a variable region encoded by a plasmid deposited with the ATCC and designated PTA-122346. In some embodiments, the TIGIT binding agent comprises a light chain encoded by a plasmid deposited with the ATCC and designated PTA-122347. In some embodiments, the TIGIT binding agent is a heavy chain comprising a variable region encoded by a plasmid deposited with the ATCC and designated PTA-122346 and a plasmid deposited with the ATCC and designated PTA-122347. Contains the encoded light chain.

In some embodiments of the methods described herein, TIGIT binding agents also include homodimeric agents/molecules and heterodimeric agents/molecules. In some embodiments, the homodimeric agent is a polypeptide. In some embodiments, the heterodimeric molecule is a polypeptide. Generally, homodimeric molecules comprise two identical polypeptides. Generally, heterodimeric molecules comprise at least two different polypeptides. In some embodiments, heterodimeric molecules, eg, bispecific agents, can bind to at least two targets. A target can be, for example, two different proteins on the surface of a single cell or two different proteins on the surface of two separate cells. The term "arm" is sometimes used herein to describe the structure of homodimeric agents, heterodimeric agents, and/or bispecific agents. In some embodiments, each arm comprises at least one polypeptide. Generally, each arm of a heterodimeric molecule has a different function, eg, binding to two different targets. In some embodiments, an arm may comprise an antigen binding site derived from an antibody. In some embodiments, one arm may comprise a binding portion of a receptor. In some embodiments, one arm may contain a ligand. In some embodiments, one arm may comprise a ligand binding region. In some embodiments, the homodimeric agent comprises two identical arms. In some embodiments, the heterodimeric agent comprises two different arms. In some embodiments, a bispecific agent comprises two different arms.

In some embodiments of the methods described herein, the invention provides TIGIT binding agents that are homodimeric molecules. In some embodiments, a homodimeric molecule comprises two identical polypeptides. In some embodiments, the invention provides TIGIT binding agents that are heterodimeric molecules. In some embodiments, the heterodimeric molecule comprises at least two different polypeptides. In some embodiments, the invention provides TIGIT binding agents that are heterodimeric agents. In some embodiments, the invention provides TIGIT binding agents that are bispecific agents. In certain embodiments, the TIGIT binding agent is a bispecific antibody.

In some aspects of the methods described herein, the invention provides polypeptides including, but not limited to, antibodies that specifically bind to TIGIT. In some embodiments, the polypeptide binds human TIGIT. In some embodiments, the polypeptide binds to mouse TIGIT. In some embodiments, the polypeptide binds to mouse TIGIT and human TIGIT. In some embodiments, the polypeptide binds to human TIGIT and not to mouse TIGIT. In some embodiments, the polypeptide binds to human TIGIT and not to rat TIGIT. In some embodiments, the polypeptide binds to human TIGIT and not to rabbit TIGIT. In some embodiments, the polypeptide binds to human TIGIT and not to marmoset TIGIT. In some embodiments, the polypeptide binds to human TIGIT and not to canine TIGIT. In some embodiments, the polypeptide binds to human TIGIT and not to porcine TIGIT. In some embodiments, the polypeptide binds to human TIGIT and not to cynomolgus monkey TIGIT. In some embodiments, the polypeptide binds to human TIGIT and not to rhesus TIGIT.

In certain embodiments of the methods described herein, the polypeptide is one, two, three, four of the CDRs of antibody OMP-313M32 (see Table 1 herein). , 5 and/or 6. In some embodiments, the polypeptide comprises CDRs with up to 4 (ie, 0, 1, 2, 3, or 4) amino acid substitutions per CDR. In certain embodiments, the heavy chain CDRs are contained within the heavy chain variable region. In certain embodiments, the light chain CDRs are contained within the light chain variable region.

In some embodiments of the methods described herein, the invention provides polypeptides that specifically bind to human TIGIT, wherein the polypeptides have at least about 80% sequence identity with SEQ ID NO:10. and/or have at least about 80% sequence identity with SEQ ID NO:11. In certain embodiments, the polypeptide has an amino acid sequence that has at least about 85%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% sequence identity with SEQ ID NO:10. including. In certain embodiments, the polypeptide is at least about 85%, at least about 90%, at least about 95%, at least about 97%, or at least about 99% the sequence of SEQ ID NO:64 or SEQ ID NO:11 Contains amino acid sequences with identity. In certain embodiments, the polypeptide has an amino acid sequence having at least about 95% sequence identity with SEQ ID NO:10 and/or an amino acid sequence having at least about 95% sequence identity with SEQ ID NO:11 including. In certain embodiments, the polypeptide comprises an amino acid sequence comprising SEQ ID NO:10 and/or an amino acid sequence comprising SEQ ID NO:11.

In some embodiments of the methods described herein, the polypeptide is SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:14 It comprises one or more amino acid sequences selected from the group consisting of NO:15, SEQ ID NO:16, and SEQ ID NO:17. Polypeptides, as defined herein, may occur as single chains or as two or more associated chains. In certain embodiments, the polypeptide comprises an amino acid sequence comprising SEQ ID NO:10 and an amino acid sequence comprising SEQ ID NO:11. In certain embodiments, the polypeptide comprises an amino acid sequence comprising SEQ ID NO:12 and an amino acid sequence comprising SEQ ID NO:14. In certain embodiments, the polypeptide comprises an amino acid sequence comprising SEQ ID NO:13 and an amino acid sequence comprising SEQ ID NO:15. In certain embodiments, the polypeptide comprises an amino acid sequence comprising SEQ ID NO:16 and an amino acid sequence comprising SEQ ID NO:14. In certain embodiments, the polypeptide comprises an amino acid sequence comprising SEQ ID NO:17 and an amino acid sequence comprising SEQ ID NO:15.

In certain embodiments of the methods described herein, the polypeptide comprises an amino acid sequence consisting of SEQ ID NO:10 and an amino acid sequence consisting of SEQ ID NO:11. In certain embodiments, the polypeptide comprises an amino acid sequence consisting of SEQ ID NO:12 and an amino acid sequence consisting of SEQ ID NO:14. In certain embodiments, the polypeptide comprises an amino acid sequence consisting of SEQ ID NO:13 and an amino acid sequence consisting of SEQ ID NO:15. In certain embodiments, the polypeptide comprises an amino acid sequence consisting of SEQ ID NO:16 and an amino acid sequence consisting of SEQ ID NO:14. In certain embodiments, the polypeptide comprises an amino acid sequence consisting of SEQ ID NO:17 and an amino acid sequence consisting of SEQ ID NO:15.

Many proteins, including antibodies, contain signal sequences that direct the transport of the protein to various locations. Generally, a signal sequence (also called a signal peptide or leader sequence) is positioned at the N-terminus of the nascent polypeptide. The signal sequence targets the polypeptide to the endoplasmic reticulum, and the protein is sorted to its destination, e.g., into the interior space of an organelle, into the inner membrane, into the outer membrane of the cell, or out of the cell via secretion. sorted. After the protein is transported to the endoplasmic reticulum, most signal sequences are cleaved from the protein by signal peptidases. Cleavage of the signal sequence from the polypeptide usually occurs at specific sites in the amino acid sequence and depends on the amino acid residues within the signal sequence. There is usually one specific cleavage site, but multiple cleavage sites may be recognized and/or used by signal peptidases to generate non-homogeneous N-termini of polypeptides. For example, using different cleavage sites within the signal sequence can result in expressed polypeptides with different N-terminal amino acids. Thus, in some embodiments, the polypeptides described herein may comprise a mixture of polypeptides with different N-termini. In some embodiments, the N-terminal length differs by 1, 2, 3, 4, or 5 amino acids. In some embodiments, the polypeptides are substantially homogeneous, ie, they have the same N-terminus. In some embodiments, the signal sequence of said polypeptide has one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.) amino acid substitutions and/or deletions. In some embodiments, the signal sequence of the polypeptide has amino acid substitutions and/or amino acid substitutions that allow a single cleavage site to predominate, thereby producing a substantially homogeneous polypeptide with an N-terminus. or contain deletions. In some embodiments, the signal sequence of the polypeptide affects the level of expression of the polypeptide, eg, increased expression or decreased expression.

を含む重鎖CDR2、および

を含む重鎖CDR3、ならびに(b)

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む、TIGIT結合物質と、TIGITとの特異的結合について競合する。 In certain embodiments of the methods described herein, the antibody or other binding agent competes with a TIGIT binding agent described herein for specific binding to human TIGIT. In some embodiments, the antibody or other binding agent comprises (a) a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

heavy chain CDR2 containing, and

a heavy chain CDR3 comprising, and (b)

Compete for specific binding to TIGIT with a TIGIT binding agent comprising a light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and a light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). do.

In certain embodiments of the methods described herein, the antibody or other binding agent comprises a heavy chain variable region comprising SEQ ID NO:10 and SEQ ID NO:11 in specific binding to human TIGIT. Compete with TIGIT binders containing light chain variable regions. In certain embodiments, the antibody or other binding agent competes for specific binding to human TIGIT with a TIGIT binding agent comprising a heavy chain comprising SEQ ID NO:13 and a light chain comprising SEQ ID NO:15. . In certain embodiments, the antibody or other binding agent competes for specific binding to human TIGIT with a TIGIT binding agent comprising a heavy chain comprising SEQ ID NO:17 and a light chain comprising SEQ ID NO:15. .

In certain embodiments of the methods described herein, the antibody or other binding agent competes with antibody OMP-313M32 for specific binding to human TIGIT. In some embodiments, the antibody or other binding agent competes with a reference antibody that is antibody OMP-313M32 for specific binding to human TIGIT. In some embodiments, the antibody or other binding agent competes with a reference antibody, which is antibody 313M33, for specific binding to human TIGIT.

In certain embodiments of the methods described herein, the antibody or other binding agent binds to the same or essentially the same epitope on TIGIT as the TIGIT binding agents described herein. In certain embodiments, the antibody or other binding agent binds to the same or essentially the same epitope on human TIGIT as antibody OMP-313M32. In certain embodiments, the antibody or other binding agent binds to the same or essentially the same epitope on human TIGIT as antibody 313M33.

In another embodiment of the methods described herein, the antibody or other binding agent binds to an epitope on TIGIT that overlaps with the epitope on TIGIT that the TIGIT binding agent described herein binds. In another embodiment, the antibody or other binding agent binds to an epitope on TIGIT that overlaps with the epitope on human TIGIT that antibody OMP-313M32 binds. In another embodiment, the antibody or other binding agent binds to an epitope on TIGIT that overlaps with the epitope on human TIGIT that antibody 313M33 binds.

In some embodiments of the methods described herein, the antibody or other binding agent competes with a TIGIT binding agent described herein for binding to an epitope comprising amino acids in SEQ ID NO:27. do. In some embodiments, the antibody or other binding agent competes with a TIGIT binding agent described herein for binding to an epitope comprising amino acids in SEQ ID NO:28. In some embodiments, the antibody or other binding agent competes with a TIGIT binding agent described herein for binding to an epitope comprising amino acids in SEQ ID NO:27 and SEQ ID NO:28. In some embodiments, the antibody or other binding agent competes with a TIGIT binding agent described herein for binding to an epitope comprising amino acids Q62 and I109 of SEQ ID NO:1. In some embodiments, the antibody or other binding agent competes with a TIGIT binding agent described herein for binding to an epitope comprising amino acids Q62 and T119 of SEQ ID NO:1. In some embodiments, the antibody or other binding agent competes with a TIGIT binding agent described herein for binding to an epitope comprising amino acids Q64 and I109 of SEQ ID NO:1. In some embodiments, the antibody or other binding agent competes with a TIGIT binding agent described herein for binding to an epitope comprising amino acids Q64 and T119 of SEQ ID NO:1. In some embodiments, the antibody or other binding agent competes with a TIGIT binding agent described herein for binding to an epitope comprising amino acids Q62, Q64, and I109 of SEQ ID NO:1. In some embodiments, the antibody or other binding agent competes with a TIGIT binding agent described herein for binding to an epitope comprising amino acids Q62, Q64, and T119 of SEQ ID NO:1. In some embodiments, the antibody or other binding agent competes with a TIGIT binding agent described herein for binding to an epitope comprising amino acids Q62, I109, and T119 of SEQ ID NO:1. In some embodiments, the antibody or other binding agent competes with a TIGIT binding agent described herein for binding to an epitope comprising amino acids Q64, I109, and T119 of SEQ ID NO:1. In some embodiments, the antibody or other binding agent competes with a TIGIT binding agent described herein for binding to an epitope comprising amino acids Q62, Q64, I109, and T119 of SEQ ID NO:1. In some embodiments, the antibody or other binding agent is selected from the group consisting of N58, E60, Q62, Q64, L65, F107, I109, H111, T117, T119, G120, and R121 of SEQ ID NO:1 Compete with the TIGIT binding agents described herein for binding to an epitope that includes at least one amino acid.

In certain embodiments of the methods described herein, the TIGIT binding agent (eg, antibody) binds to TIGIT and modulates TIGIT activity. In some embodiments, the TIGIT binding agent is a TIGIT antagonist and reduces TIGIT activity. In certain embodiments, the TIGIT binding agent inhibits TIGIT activity by at least about 10%, at least about 20%, at least about 30%, at least about 50%, at least about 75%, at least about 90%, or about 100%. In certain embodiments, the TIGIT binding agent that inhibits TIGIT activity is the antibody OMP-313M32. In certain embodiments, the TIGIT binding agent that inhibits TIGIT activity is the antibody OMP-313M32. In certain embodiments, the TIGIT binding agent that inhibits TIGIT activity is the antibody OMP-313M33.

In some embodiments of the methods described herein, the TIGIT binding agent binds to TIGIT and inhibits or reduces TIGIT signaling. In certain embodiments, the TIGIT binding agent (e.g., antibody) inhibits TIGIT signaling by at least about 10%, at least about 20%, at least about 30%, at least about 50%, at least about 75%, at least about 90%, or Inhibits approximately 100%. In some embodiments, the TIGIT binding agent inhibits mouse TIGIT signaling. In some embodiments, the TIGIT binding agent inhibits human TIGIT signaling. In certain embodiments, the TIGIT binding agent that inhibits TIGIT signaling is the antibody OMP-313M32. In certain embodiments, the TIGIT binding agent that inhibits TIGIT signaling is the antibody OMP-313M33.

After TIGIT interacts with the TIGIT counter-receptor PVR, the cytoplasmic tail of TIGIT is phosphorylated. TIGIT phosphorylation is the beginning of a cascade involving downstream events that affect other known signaling pathways. Therefore, assessing TIGIT phosphorylation can provide information about TIGIT activity and TIGIT signaling.

Phosphorylation assays are known to those of skill in the art and are commonly used to monitor protein activation and/or pathway activation. This assay may be used to monitor the effects of various treatments on target protein and/or target pathway activation. For example, an in vitro phosphorylation assay can be used to assess the effect of a TIGIT antagonist on PVR-induced TIGIT activation.

In certain embodiments of the methods described herein, the TIGIT binding agent (eg, antibody) inhibits binding of TIGIT to the receptor. In certain embodiments, the TIGIT binding agent inhibits binding between TIGIT and PVR. In some embodiments, a TIGIT binding agent inhibits binding of TIGIT to PVR-L2, PVR-L3, and/or PVR-L4. In certain embodiments, the inhibition of binding of a TIGIT binding agent to PVR is at least about 10%, at least about 25%, at least about 50%, at least about 75%, at least about 90%, or at least about 95%. . In certain embodiments, the inhibition of binding of a TIGIT binding agent to PVR-L2, PVR-L3, and/or PVR-L4 is at least about 10%, at least about 25%, at least about 50%, at least about 75% , at least about 90%, or at least about 95%. In certain embodiments, the TIGIT-binding agent that inhibits binding of TIGIT to PVR is the antibody OMP-313M32. In certain embodiments, the TIGIT-binding agent that inhibits binding of TIGIT to PVR is the antibody OMP-313M33. In certain embodiments, a TIGIT-binding agent that inhibits binding of TIGIT to PVR-L2, PVR-L3, and/or PVR-L4 is the antibody OMP-313M32. In certain embodiments, a TIGIT-binding agent that inhibits binding of TIGIT to PVR-L2, PVR-L3, and/or PVR-L4 is antibody OMP-313M33.

In certain embodiments of the methods described herein, the TIGIT binding agent (eg, antibody) blocks binding of TIGIT to the receptor. In certain embodiments, the TIGIT binding agent blocks binding between TIGIT and PVR. In certain embodiments, the blocking of binding between a TIGIT binding agent and a PVR is at least about 10%, at least about 25%, at least about 50%, at least about 75%, at least about 90%, or at least about 95%. . In some embodiments, the TIGIT binding agent blocks binding of TIGIT to PVRL2, PVRL3, and/or PVRL4. In certain embodiments, the blocking of binding of a TIGIT binding agent to PVRL2, PVRL3, and/or PVRL4 is at least about 10%, at least about 25%, at least about 50%, at least about 75%, at least about 90%, or at least about 95%. In certain embodiments, the TIGIT binding agent that blocks binding of TIGIT to PVR is the antibody OMP-313M32. In certain embodiments, the TIGIT binding agent that blocks binding of TIGIT to PVR is the antibody OMP-313M33. In certain embodiments, a TIGIT-binding agent that blocks binding of TIGIT to PVRL2, PVRL3, and/or PVRL4 is the antibody OMP-313M32. In certain embodiments, a TIGIT-binding agent that blocks binding of TIGIT to PVRL2, PVRL3, and/or PVRL4 is the antibody OMP-313M33.

Binding assays are known to those of skill in the art and are described herein. Binding assays may be used to monitor the effect of a test agent on the interaction of a target protein with its binding partner (eg, receptor or ligand). For example, an in vitro binding assay can be used to assess whether a TIGIT antagonist blocks the interaction of TIGIT and PVR.

In certain embodiments, the TIGIT-binding agents described herein have the following effects: inhibit tumor cell proliferation; inhibit tumor growth; reducing the tumorigenicity of a tumor by reducing the frequency of cancer stem cells, inducing cell death of tumor cells, enhancing or boosting an immune response, enhancing or boosting an anti-tumor response, immunity increasing the cytolytic activity of cells, increasing killing of tumor cells, increasing killing of tumor cells by immune cells, inducing cells within tumors to differentiate, transforming tumorigenic cells into non-tumor cells differentiating to a formed state, inducing expression of differentiation markers in tumor cells, inhibiting metastasis of tumor cells, decreasing survival of tumor cells, increasing inhibition of cell contact dependent growth, tumors It has one or more of increasing cell apoptosis, decreasing epithelial-mesenchymal transition (EMT), or decreasing tumor cell survival. In some embodiments, the agent has one or more of the following effects: inhibiting viral infection, inhibiting chronic viral infection, reducing viral load, reducing virus-infected cells. Inducing death or reducing the number or percentage of virus-infected cells.

In certain embodiments of the methods described herein, the TIGIT binding agent inhibits tumor growth. In certain embodiments, TIGIT binding agents inhibit tumor growth in vivo (eg, in mouse models and/or in humans with cancer). In certain embodiments, tumor growth is inhibited at least about 2-fold, about 3-fold, about 5-fold, about 10-fold, about 50-fold, about 100-fold, or about 1000-fold compared to untreated tumors.

In certain embodiments of the methods described herein, agents (eg, polypeptides and/or antibodies) bind TIGIT and modulate an immune response. In some embodiments, a TIGIT binding agent activates and/or increases an immune response. In some embodiments, the TIGIT binding agent increases, promotes, or enhances cell-mediated immunity. In some embodiments, the TIGIT binding agent increases, promotes, or enhances innate cell-mediated immunity. In some embodiments, the TIGIT binding agent increases, promotes, or enhances adaptive cell-mediated immunity. In some embodiments, the TIGIT binding agent increases, promotes, or enhances T cell activity. In some embodiments, the TIGIT binding agent increases, promotes, or enhances cytolytic T cell (CTL) activity. In some embodiments, the TIGIT binding agent increases, promotes, or enhances NK cell activity. In some embodiments, the TIGIT binding agent increases, promotes, or enhances lymphokine-activated killer cell (LAK) activity. In some embodiments, the TIGIT-binding agent increases, promotes, or enhances tumor infiltrating lymphocyte (TIL) activity. In some embodiments, the TIGIT binding agent inhibits or reduces Treg cell activity. In some embodiments, the TIGIT binding agent inhibits or reduces MDSC activity. In some embodiments, the TIGIT binding agent increases, promotes, or enhances tumor cell killing. In some embodiments, the TIGIT binding agent increases, promotes, or enhances inhibition of tumor growth.

In certain embodiments of the methods described herein, the TIGIT binding agent is an antagonist of human TIGIT. In some embodiments, the agent is an antagonist of TIGIT and activates and/or increases an immune response. In some embodiments, the agent is an antagonist of TIGIT and activates and/or increases the activity of NK cells. In certain embodiments, the agent increases the activity by at least about 10%, at least about 20%, at least about 30%, at least about 50%, at least about 75%, at least about 90%, or about 100%. . In some embodiments, the agent is an antagonist of TIGIT and activates and/or increases T cell activity (eg, T cell cytolytic activity). In certain embodiments, said agent increases said activity by at least about 10%, at least about 20%, at least about 30%, at least about 50%, at least about 75%, at least about 90%, or about 100%. In some embodiments, the agent is an antagonist of TIGIT and induces and/or enhances a Th1-type immune response. In general, Th1-type immune responses involve production of interferon-γ (IFN-γ), IL-2, and tumor necrosis factor-β (TNF-β). In comparison, Th2-type immune responses generally involve production of IL-4, IL-5, IL-6, IL-9, IL-10, and IL-13. In some embodiments, the agent is an antagonist of TIGIT and induces and/or increases cytokine or lymphokine production. In some embodiments, the induction and/or enhancement of cytokine or lymphokine production may be an indirect effect.

In certain embodiments of the methods described herein, the TIGIT binding agent increases NK cell activation. In certain embodiments, the TIGIT binding agent increases T cell activation. In certain embodiments, activation of NK cells and/or T cells by a TIGIT binding agent reduces the level of NK cell and/or T cell activation by at least about 10%, at least about 25%, at least about 50%. %, at least about 75%, at least about 90%, or at least about 95%. In certain embodiments, the TIGIT binding agent that increases NK cell activation is the antibody OMP-313M32. In certain embodiments, the TIGIT binding agent that increases NK cell activation is the antibody OMP-313M33.

In certain embodiments of the methods described herein, the TIGIT binding agent (eg, antibody) is an antagonist of regulatory T cell (Treg) activity. In certain embodiments, the TIGIT binding agent inhibits or reduces Treg activity. In certain embodiments, inhibition of Treg activity by a TIGIT binding agent reduces the suppressive activity of Treg cells by at least about 10%, at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least About 95% or about 100% inhibition. In certain embodiments, the TIGIT binding agent that inhibits Treg activity is the antibody OMP-313M32. In certain embodiments, the TIGIT binding agent that inhibits Treg activity is the antibody OMP-313M33.

In certain embodiments of the methods described herein, the TIGIT-binding agent (eg, antibody) is an antagonist of myeloid-derived suppressor cells (MDSCs). In certain embodiments, the TIGIT binding agent inhibits MDSC activity. In certain embodiments, the TIGIT binding agent inhibits MDSC activity by at least about 10%, at least about 20%, at least about 30%, at least about 50%, at least about 75%, at least about 90%, or about 100%. In certain embodiments, the TIGIT binding agent that inhibits MDSC activity is the antibody OMP-313M32. In certain embodiments, the TIGIT binding agent that inhibits MDSC activity is the antibody OMP-313M33.

In certain embodiments of the methods described herein, the TIGIT binding agent (eg, antibody) increases natural killer (NK) cell activity. In certain embodiments, the TIGIT binding agent increases NK cell activity by at least about 10%, at least about 20%, at least about 30%, at least about 50%, at least about 75%, at least about 90%, or about 100%. . In certain embodiments, the TIGIT binding agent that increases NK cell activity is the antibody OMP-313M32. In certain embodiments, the TIGIT binding agent that increases NK cell activity is the antibody OMP-313M33.

In certain embodiments of the methods described herein, the TIGIT binding agent (eg, antibody) increases tumor infiltrating lymphocyte (TIL) activity. In certain embodiments, the TIGIT binding agent increases TIL activity by at least about 10%, at least about 20%, at least about 30%, at least about 50%, at least about 75%, at least about 90%, or about 100%. In certain embodiments, the TIGIT binding agent that increases TIL cell activity is the antibody OMP-313M32. In certain embodiments, the TIGIT binding agent that increases TIL cell activity is the antibody OMP-313M33.

In certain embodiments of the methods described herein, the TIGIT binding agent (eg, antibody) increases or enhances lymphokine-activated killer cell (LAK) activity. In certain embodiments, the TIGIT binding agent increases LAK activity by at least about 10%, at least about 20%, at least about 30%, at least about 50%, at least about 75%, at least about 90%, or about 100%. In certain embodiments, the TIGIT binding agent that increases LAK cell activity is the antibody OMP-313M32. In certain embodiments, the TIGIT binding agent that increases LAK cell activity is the antibody OMP-313M33.

In vivo and in vitro assays for determining whether a TIGIT binding agent (or candidate binding agent) modulates an immune response are known in the art or are being developed. In some embodiments, functional assays that detect T cell activation can be used. In some embodiments, functional assays that detect T cell proliferation can be used. In some embodiments, functional assays that detect NK activity can be used. In some embodiments, functional assays that detect CTL activity can be used. In some embodiments, functional assays that detect Treg activity can be used. In some embodiments, functional assays that detect MDSC activity can be used. In some aspects, functional assays that detect cytokine or lymphokine production, or cells that produce cytokines or lymphokines, can be used. In some embodiments, an ELISPOT assay is used to measure the frequency of antigen-specific T cells. In some embodiments, ELISPOT assays are used to measure cytokine release/production and/or are used to measure the number of cells that produce cytokines. In some embodiments, cytokine assays are used to identify Th1-type responses. In some embodiments, cytokine assays are used to identify Th2-type responses. In some embodiments, cytokine assays are used to identify Th17-type responses. In some embodiments, FACS analysis is used to measure activation markers on immune cells, including but not limited to T cells, B cells, NK cells, macrophages, and/or myeloid cells.

In certain embodiments of the methods described herein, the TIGIT binding agent is administered for at least about 2 hours, at least about 5 hours, at least about 10 hours, at least about 24 hours, at least about 3 days, at least about 1 week, or It has a circulation half-life in mice, rats, cynomolgus monkeys, or humans of at least about 2 weeks. In certain embodiments, the TIGIT-binding agent is administered to mice, cynomolgus monkeys for at least about 2 hours, at least about 5 hours, at least about 10 hours, at least about 24 hours, at least about 3 days, at least about 1 week, or at least about 2 weeks. , or an IgG (eg, IgG1, IgG2, or IgG4) antibody that has a circulating half-life in humans. Methods to extend (or shorten) the half-life of agents such as polypeptides and antibodies are known in the art. For example, known methods of increasing the circulation half-life of IgG antibodies include introducing mutations into the Fc region that increase pH-dependent binding of the antibody to fetal Fc receptors (FcRn) at pH 6.0. . Known methods of increasing the circulation half-life of antibody fragments lacking the Fc region include techniques such as pegylation.

In some embodiments of the methods described herein, the TIGIT binding agent is a polypeptide. In some embodiments, the polypeptide is a recombinant, natural, or synthetic polypeptide comprising an antibody or fragment thereof that binds TIGIT. It is recognized in the art that some amino acid sequences of the present invention can be altered without significantly affecting protein structure or function. Accordingly, the invention further includes variations of said polypeptides that exhibit considerable activity in binding to TIGIT or that comprise regions of antibodies or fragments thereof that bind to TIGIT. In some embodiments, amino acid sequence variations of TIGIT-binding polypeptides include deletions, insertions, inversions, repeats, and/or other types of substitutions.

The polypeptides, analogs and variants thereof can be further modified to contain additional chemical moieties not normally a part of the polypeptide. Derivatizing moieties can improve or otherwise modulate the solubility, biological half-life, and/or absorption of the polypeptide. These moieties can also reduce or eliminate unwanted side effects of the polypeptides and variants. A summary of chemical moieties can be found in Remington: The Science and Practice of Pharmacy, 22nd ^Edition , 2012, Pharmaceutical Press, London.

In certain embodiments of the methods described herein, the TIGIT binding agent is in any one of a number of conjugated forms (i.e., immunoconjugates or radioconjugates) or unconjugated forms. Used. In certain embodiments, the agent uses the subject's natural defense mechanisms, including complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC), to eliminate malignant or cancer cells. It can be used in unconjugated form.

In some embodiments of the methods described herein, the TIGIT binding agent is conjugated to a cytotoxic agent. In some embodiments, the TIGIT binding agent is an antibody conjugated to a cytotoxic agent as an ADC (antibody-drug conjugate). In some embodiments, cytotoxic agents include, but are not limited to, methotrexate, adriamycin/doxorubicin, melphalan, mitomycin C, chlorambucil, daunorubicin, pyrrolobenzodiazepine (PBD), or other intercalating agents. is. In some embodiments, the cytotoxic agent is a toxin with enzymatic activity derived from bacteria, fungi, plants, or animals, or diphtheria A chain, non-binding active fragments of diphtheria toxin, exotoxin A chain, ricin A chain. , abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii protein, diancin protein, Phytolaca americana protein (PAPI, PAPII, and PAP-S), bitter melon (Momordica charantia) inhibitor , curcin, crotin, Sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, and tricothecene, which Fragments. In some embodiments, the cytotoxic agent is a radioisotope to generate radioconjugates or radioconjugate antibodies. ⁹⁰ Y, ¹²⁵ I, ¹³¹ I, ¹²³ I, ¹¹¹ In, ¹³¹ In, ¹⁰⁵ Rh, ¹⁵³ Sm, ⁶⁷ Cu, ⁶⁷ Ga, ¹⁶⁶ Ho, ¹⁷⁷ Lu, ¹⁸⁶ Re, ¹⁸⁸ Re were used to generate radioconjugated antibodies. , and ²¹² Bi can be used. Conjugates of antibodies with one or more small molecule toxins such as calicheamicin, maytansinoids, trichothenes, and CC1065, and derivatives of these toxins that have toxin activity can also be used. . Conjugates of antibodies and cytotoxic agents can be made with a variety of bifunctional protein coupling agents such as N-succinimidyl-3-(2-pyridyidithiol) propionate (SPDP), iminothiolane (IT), Bifunctional derivatives of imidoesters (e.g. dimethyl adipimidate HCl), active esters (e.g. disuccinimidyl suberate), aldehydes (e.g. glutaraldehyde), bis-azido compounds (e.g. bis(p-azidobenzoyl )hexanediamine), bis-diazonium derivatives (e.g. bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (e.g. toluene 2,6-diisocyanate), and bis-active fluorine compounds (e.g. 1,5-difluoro- 2,4-dinitrobenzene).

IV. Polynucleotides The present invention provides polynucleotides, including polynucleotides that encode TIGIT binding agents. The term "polynucleotide encoding a polypeptide" encompasses polynucleotides that contain only coding sequences for the polypeptide, as well as polynucleotides that contain additional coding and/or non-coding sequences. A polynucleotide of the invention may be in the form of RNA or it may be in the form of DNA. DNA includes cDNA, genomic DNA, and synthetic DNA, and may be double-stranded or single-stranded, and if single stranded may be the coding strand or non-coding (anti-sense) strand.

In certain embodiments, the polynucleotide comprises SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO: 16, and SEQ ID NO:17.

In certain embodiments, the polynucleotide comprises SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO: 16, and SEQ ID NO:17, at least about 80% identical, at least about 85% identical, at least about 90% identical, at least about 95% identical to a polynucleotide encoding an amino acid sequence selected from the group consisting of , in some embodiments, comprise polynucleotides having nucleotide sequences that are at least about 96%, 97%, 98%, or 99% identical. consisting of SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, and SEQ ID NO:17 Polynucleotides comprising polynucleotides that hybridize to polynucleotides encoding amino acid sequences selected from the group are also provided. In certain embodiments, hybridization is performed under high stringency conditions. Conditions of high stringency are known to those skilled in the art and include: (1) low ionic strength and high temperature for washing, e.g., 15 mM sodium chloride/1.5 mM sodium citrate (1×SSC) and 0.1 conditions using % sodium dodecyl sulfate; (2) denaturants, such as formamide, dissolved in 5×SSC (0.75 M NaCl, 75 mM sodium citrate) during hybridization; 50% (v/v); conditions using formamide and 0.1% bovine serum albumin/0.1% Ficoll/0.1% polyvinylpyrrolidone/50 mM sodium phosphate buffer, pH 6.5 at 42°C; or (3) 50% formamide, 5 x SSC, 50 mM phosphorus. sodium sulfate (pH 6.8), 0.1% sodium pyrophosphate, 5x Denhardt's solution, sonicated salmon sperm DNA (50 µg/ml), 0.1% SDS, and 10% dextran sulfate at 42°C and 55°C. 0.2×SSC containing 50% formamide at 55° C., followed by a high stringency wash consisting of 0.1×SSC containing EDTA at 55° C., but not limited thereto.

In some embodiments, the polynucleotide is a polynucleotide sequence selected from the group consisting of SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22 including. In certain embodiments, the polynucleotide comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22. at least about 80% identical, at least about 85% identical, at least about 90% identical, at least about 95% identical, in some embodiments at least about 96%, 97%, 98%, or 99% identical It includes a polynucleotide having a nucleotide sequence. A polynucleotide comprising a polynucleotide that hybridizes to a polynucleotide sequence selected from the group consisting of SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, and SEQ ID NO:22 is also provided. In certain embodiments, hybridization methods are performed under high stringency conditions as described above.

In certain embodiments, the polynucleotide is in the same reading frame, e.g., a polynucleotide that aids in the expression and secretion of a polypeptide from a host cell (e.g., a secretory agent to control transport of a polypeptide out of a cell). It contains the coding sequence for the mature polypeptide fused with a leader sequence that functions as a sequence. A polypeptide with a leader sequence is a preprotein and may have a leader sequence that is cleaved by the host cell to form the mature polypeptide. The polynucleotide may also encode a proprotein that is the mature protein plus additional 5' amino acid residues. A mature protein with a prosequence is a proprotein and is an inactive protein. Cleavage of the prosequence leaves an active, mature protein.

In certain embodiments, a polynucleotide comprises a mature polypeptide coding sequence fused, in the same reading frame, to a marker sequence, eg, a marker sequence that allows purification of the encoded polypeptide. For example, the marker sequence can be a hexahistidine tag supplied by the pQE-9 vector for purification of the mature polypeptide fused to the marker in the case of bacterial hosts. Alternatively, the marker sequence may be a hemagglutinin (HA) tag derived from the influenza hemagglutinin protein when mammalian hosts (eg, COS-7 cells) are used. In some embodiments, the marker sequence is the FLAG-tag, which can be used with other affinity tags and is a peptide of sequence DYKDDDDK (SEQ ID NO:29).

The invention further relates to variants of the polynucleotides described herein, said variants encoding, for example, fragments, analogues and/or derivatives.

In certain embodiments, the present invention is at least about 80% identical, at least about 85% identical, at least about 90% identical, at least about 95% identical, at least about 95% identical, to a polynucleotide encoding a polypeptide comprising a TIGIT binding agent. Embodiments provide polynucleotides, including polynucleotides having nucleotide sequences that are at least about 96%, 97%, 98%, or 99% identical.

As used herein, the phrase a polynucleotide having a nucleotide sequence that is at least, e.g., 95% "identical" to a reference nucleotide sequence means that said polynucleotide sequence is at up to 5 points of every 100 nucleotides of the reference nucleotide sequence. It is intended to mean that the nucleotide sequence of said polynucleotide is identical to the reference sequence, except that it may contain mutations. In other words, up to 5% of the nucleotides in the reference sequence can be deleted or replaced with different nucleotides to obtain a polynucleotide having a nucleotide sequence that is at least 95% identical to the reference nucleotide sequence. Alternatively, up to 5% of the total nucleotides in the reference sequence can be inserted into the reference sequence. These mutations of the reference sequence may occur at the 5' or 3' terminal position of the reference nucleotide sequence or anywhere between the terminal positions and may be individually interspersed between nucleotides in the reference sequence. They may be interspersed in one or more contiguous groups within the sequence.

A polynucleotide variant may contain alterations in the coding regions, non-coding regions, or both. In some embodiments, polynucleotide variants contain changes that produce silent substitutions, additions, or deletions, but do not alter the properties or activities of the encoded polypeptide. In some embodiments, the polynucleotide variant contains silent substitutions that do not alter the amino acid sequence of the polypeptide (due to the degeneracy of the genetic code). Polynucleotide variants can be generated for a variety of reasons, e.g., to optimize codon expression for a particular host (i.e., change the codons of human mRNA to codons preferred by a bacterial host, e.g., E. coli). can change). In some embodiments, the polynucleotide variant contains at least one silent mutation in the noncoding or coding region of the sequence.

In some embodiments, polynucleotide variants are generated to modulate or alter the expression (or level of expression) of the encoded polypeptide. In some embodiments, polynucleotide variants are generated to increase expression of the encoded polypeptide. In some embodiments, polynucleotide variants are generated to reduce expression of the encoded polypeptide. In some embodiments, the polynucleotide variant increases expression of the encoded polypeptide compared to the parent polynucleotide sequence. In some embodiments, the polynucleotide variant has reduced expression of the encoded polypeptide compared to the parent polynucleotide sequence.

In some embodiments, at least one polynucleotide variant is generated (without altering the amino acid sequence of the encoded polypeptide) to increase production of heterodimeric molecules. In some embodiments, at least one polynucleotide variant is generated (without altering the amino acid sequence of the encoded polypeptide) to increase production of bispecific antibodies.

In certain embodiments, said polynucleotide is isolated. In certain embodiments, said polynucleotide is substantially pure.

Vectors and cells comprising the polynucleotides described herein are also provided. In some embodiments, the expression vector comprises a polynucleotide molecule. In some embodiments, the host cell comprises an expression vector comprising said polynucleotide molecule. In some embodiments, the host cell contains the polynucleotide molecule.

V. Kits Containing Agents Described Herein The invention provides kits containing TIGIT binding agents described herein that can be used to practice the methods described herein. . In certain embodiments, kits comprise at least one purified TIGIT binding agent in one or more containers. In some embodiments, the kit includes components necessary and/or sufficient to perform a detection assay, including all controls, instructions for performing the assay, and any necessary components for analyzing and presenting the results. Includes all software. Those skilled in the art will readily recognize that the disclosed TIGIT binding agents of the present invention can be readily incorporated into one of the established kit formats well known in the art.

Further provided are kits comprising a TIGIT binding agent and at least one additional therapeutic agent. In certain embodiments, the second (or third or more) therapeutic agent is a chemotherapeutic agent. In certain embodiments, the second (or third or more) therapeutic agent is an antibody.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、高頻度MSI腫瘍は、CRC、胃がん、子宮内膜がん、肝胆道がん、尿路がん、脳がん、および皮膚がんからなる群より選択される。一部の態様において、TIGITの細胞外ドメインに特異的に結合する抗体は、約0.3mg/kg、約1mg/kg、約3mg/kg、約10mg/kg、または約15mg/kgの用量で2週間ごとに投与される。 VI. Further Exemplary Embodiments In some embodiments, the methods of inhibiting tumor growth in a subject provided throughout this application are directed to a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of TIGIT. wherein the tumor is a high frequency MSI solid tumor and the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the frequent MSI tumor is selected from the group consisting of CRC, stomach cancer, endometrial cancer, hepatobiliary cancer, urinary tract cancer, brain cancer, and skin cancer. In some embodiments, an antibody that specifically binds to the extracellular domain of TIGIT is administered at a dose of about 0.3 mg/kg, about 1 mg/kg, about 3 mg/kg, about 10 mg/kg, or about 15 mg/kg. Administered weekly.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments, the method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of TIGIT, wherein the tumor is MSI CRC, MSI gastric cancer , MSI endometrial cancer, MSI hepatobiliary cancer, MSI urinary tract cancer, MSI brain cancer, and MSI skin cancer, wherein the antibody is about 3 mg /kg every two weeks and antibodies that bind to human TIGIT are heavy chain CDR1s containing TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments, the method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of TIGIT, wherein the tumor is MSI CRC, MSI gastric cancer , MSI endometrial cancer, MSI hepatobiliary cancer, MSI urinary tract cancer, MSI brain cancer, and MSI skin cancer, wherein the antibody is about 10 mg /kg every two weeks and antibodies that bind to human TIGIT are heavy chain CDR1s containing TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

, light chain CDR2 with SASYRYT (SEQ ID NO:8), and light chain CDR3 with QQHYSTP (SEQ ID NO:9).

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments, the method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of TIGIT, wherein the tumor is MSI CRC, MSI gastric cancer , MSI endometrial cancer, MSI hepatobiliary cancer, MSI urinary tract cancer, MSI brain cancer, and MSI skin cancer, wherein the antibody is administered for 2 weeks The antibody that binds to human TIGIT is administered at a dose of about 15 mg/kg per day, and that binds to human TIGIT is a heavy chain CDR1 containing TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、TIGITの細胞外ドメインに特異的に結合する抗体は、約0.3mg/kg、約1mg/kg、約3mg/kg、約10mg/kg、または約15mg/kgの用量で2週間ごとに投与される。 In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of TIGIT, wherein the tumor is melanoma, NSCLC, an anti-PD1 antibody selected from the group consisting of renal cell carcinoma, head and neck squamous cell carcinoma, urothelial carcinoma, colorectal cancer (e.g., MSI or dMMR metastatic CRC), and hepatocellular carcinoma Antibodies that are tumor refractory to treatment with and that bind to human TIGIT are heavy chain CDR1s containing TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, an antibody that specifically binds to the extracellular domain of TIGIT is administered at a dose of about 0.3 mg/kg, about 1 mg/kg, about 3 mg/kg, about 10 mg/kg, or about 15 mg/kg. Dosed weekly.

In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of TIGIT, wherein the tumor is melanoma, NSCLC, anti-PD-L1 selected from the group consisting of renal cell carcinoma, head and neck squamous cell carcinoma, urothelial carcinoma, colorectal cancer (e.g., MSI or dMMR metastatic CRC), and hepatocellular carcinoma Antibodies that are tumors refractory to treatment with antibodies and that bind to human TIGIT have heavy chain CDR1 containing TSDYAWN (SEQ ID NO:4), heavy chain CDR2 containing YISYSGSTSYNPSLRS (SEQ ID NO:5) , heavy chain CDR3 with ARRQVGLGFAY (SEQ ID NO:6), light chain CDR1 with KASQDVSTAVA (SEQ ID NO:7), light chain CDR2 with SASYRYT (SEQ ID NO:8), and QQHYSTP (SEQ ID NO: 9) containing the light chain CDR3. In some embodiments, an antibody that specifically binds to the extracellular domain of TIGIT is administered at a dose of about 0.3 mg/kg, about 1 mg/kg, about 3 mg/kg, about 10 mg/kg, or about 15 mg/kg. Administered weekly.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、対象は抗PD1抗体を用いて以前に処置されたことがある。一部の態様において、TIGITの細胞外ドメインに特異的に結合する抗体は、約0.3mg/kg、約1mg/kg、約3mg/kg、約10mg/kg、または約15mg/kgの用量で2週間ごとに投与される。 In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of TIGIT, wherein the tumor is melanoma, NSCLC, an anti-PD1 antibody selected from the group consisting of renal cell carcinoma, head and neck squamous cell carcinoma, urothelial carcinoma, colorectal cancer (e.g., MSI or dMMR metastatic CRC), and hepatocellular carcinoma Antibodies that are tumors refractory to treatment with and that bind to human TIGIT are heavy chain CDR1s containing TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the subject has been previously treated with an anti-PD1 antibody. In some embodiments, an antibody that specifically binds to the extracellular domain of TIGIT is administered at a dose of about 0.3 mg/kg, about 1 mg/kg, about 3 mg/kg, about 10 mg/kg, or about 15 mg/kg. Dosed weekly.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、対象は抗PD-L1抗体を用いて以前に処置されたことがある。一部の態様において、TIGITの細胞外ドメインに特異的に結合する抗体は、約0.3mg/kg、約1mg/kg、約3mg/kg、約10mg/kg、または約15mg/kgの用量で2週間ごとに投与される。 In some embodiments, a method of inhibiting tumor growth in a subject comprises administering to the subject a therapeutically effective amount of an antibody that specifically binds to the extracellular domain of TIGIT, wherein the tumor is melanoma, NSCLC, anti-PD-L1 selected from the group consisting of renal cell carcinoma, head and neck squamous cell carcinoma, urothelial carcinoma, colorectal cancer (e.g., MSI or dMMR metastatic CRC), and hepatocellular carcinoma Antibodies that are tumors that are refractory to treatment with antibodies and that bind to human TIGIT are heavy chain CDR1s containing TSDYAWN (SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the subject has been previously treated with an anti-PD-L1 antibody. In some embodiments, an antibody that specifically binds to the extracellular domain of TIGIT is administered at a dose of about 0.3 mg/kg, about 1 mg/kg, about 3 mg/kg, about 10 mg/kg, or about 15 mg/kg. Dosed weekly.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。一部の態様において、前記腫瘍は、頭頸部がん、食道がん、胃がん、子宮頸がん、TNBR、肛門がん、肝細胞がん、高頻度MSI固形腫瘍(例えば、MSI CRC)、NSCLC、および結腸直腸がんからなる群より選択される。一部の態様において、TIGIの細胞外ドメインに特異的に結合する抗体は、約0.3mg/kg、約1mg/kg、約3mg/kg、約10mg/kg、または約15mg/kgの用量で2週間ごとに投与される。 In some embodiments, the method of inhibiting tumor growth in a subject is based on the tumor/cancer expressing PVR and/or PVRL2, a therapeutically effective amount of selectively administering an antibody to the subject, wherein the antibody that binds to human TIGIT is a heavy chain CDR1 comprising TSDYAWN (SEQ ID NO:4);

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9). In some embodiments, the tumor is head and neck cancer, esophageal cancer, gastric cancer, cervical cancer, TNBR, anal cancer, hepatocellular carcinoma, MSI frequent solid tumors (eg, MSI CRC), NSCLC , and colorectal cancer. In some embodiments, the antibody that specifically binds to the extracellular domain of TIGI is administered at a dose of about 0.3 mg/kg, about 1 mg/kg, about 3 mg/kg, about 10 mg/kg, or about 15 mg/kg. Administered weekly.

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments, the method of inhibiting tumor growth in a subject is based on the tumor/cancer expressing PVR and/or PVRL2, a therapeutically effective amount of Tumor/cancer includes head and neck cancer, esophageal cancer, gastric cancer, cervical cancer, TNBR, anal cancer, hepatocellular carcinoma, high frequency MSI solid The antibody is selected from the group consisting of tumors (e.g., MSI CRC), NSCLC, and colorectal cancer, wherein the antibody is administered at a dose of about 3 mg/kg every two weeks, and the antibody that binds human TIGIT is TSDYAWN ( heavy chain CDR1 comprising SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments, the method of inhibiting tumor growth in a subject is based on the tumor/cancer expressing PVR and/or PVRL2, a therapeutically effective amount of Tumor/cancer includes head and neck cancer, esophageal cancer, gastric cancer, cervical cancer, TNBR, anal cancer, hepatocellular carcinoma, high frequency MSI solid The antibody is selected from the group consisting of tumor (e.g., MSI CRC), NSCLC, and colorectal cancer, wherein said antibody is administered at a dose of about 10 mg/kg every two weeks, and the antibody that binds to human TIGIT is TSDYAWN ( heavy chain CDR1 comprising SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

を含む重鎖CDR2、

を含む重鎖CDR3、

を含む軽鎖CDR1、SASYRYT(SEQ ID NO:8)を含む軽鎖CDR2、およびQQHYSTP(SEQ ID NO:9)を含む軽鎖CDR3を含む。 In some embodiments, the method of inhibiting tumor growth in a subject is based on the tumor/cancer expressing PVR and/or PVRL2, a therapeutically effective amount of Tumor/cancer includes head and neck cancer, esophageal cancer, gastric cancer, cervical cancer, TNBR, anal cancer, hepatocellular carcinoma, high frequency MSI solid The antibody is selected from the group consisting of tumor (e.g., MSI CRC), NSCLC, and colorectal cancer, wherein said antibody is administered at a dose of about 15 mg/kg every two weeks, and the antibody that binds to human TIGIT is TSDYAWN ( heavy chain CDR1 comprising SEQ ID NO:4),

a heavy chain CDR2 comprising

a heavy chain CDR3 comprising

light chain CDR1 comprising SASYRYT (SEQ ID NO:8), and light chain CDR3 comprising QQHYSTP (SEQ ID NO:9).

Aspects of the present disclosure can be further clarified by reference to the following non-limiting examples. The following non-limiting examples detail the preparation of certain antibodies of this disclosure and methods of using the antibodies of this disclosure. It will be apparent to those skilled in the art that many modifications, both to materials and methods, can be practiced without departing from the scope of this disclosure.

Example 1
In Vivo Tumor Growth Inhibition in Humanized Mice by Anti-TIGIT Antibodies A humanized mouse model was used to test the efficacy of treatment with anti-hTIGIT antibodies against human tumors. Humanized mice were obtained from Jackson Laboratories. These mice were generated by injecting human hematopoietic stem cells (CD34+ cells) into irradiated NSG mice. After 15 weeks, the presence of mature human lymphocytes was confirmed by flow cytometry. Each mouse was injected subcutaneously with patient-derived melanoma tumor cells (OMP-M9; 75,000 cells/mouse). Tumors were grown for 19 days until the average tumor volume was approximately 50 mm ³ . Tumor-bearing mice were randomized into groups (n=8 mice/group). Tumor-bearing mice were treated with control antibody or anti-hTIGIT antibody OMP-313M32. Mice were dosed with 1 mg/kg or 5 mg/kg every 5 days. Tumor growth was monitored and tumor volumes were measured using electronic calipers at the indicated time points.

As shown in Figure 1, tumor growth was inhibited in mice treated with antibody OMP-313M32 compared to controls. These results indicate that TIGIT targeting was effective in augmenting anti-tumor immune responses of human lymphocytes and contributed to human tumor growth inhibition in vivo. Furthermore, in parallel with preclinical studies performed with anti-TIGIT antibodies 313R12 and 313R19 in mouse tumor models, these results suggest that humanized mouse models with patient-derived xenografts (which bind only to human TIGIT ), it proved possible to study the anti-hTIGIT antibody OMP-313M32.

Example 2
Phase 1 Study An open-label Phase 1 dose escalation and escalation study of OMP-313M32 in patients with certain locally advanced or metastatic tumors was designed. Patients have tumors that have progressed after standard therapy or have tumors that have proven intolerable or deemed inappropriate for therapy. Prior to enrollment, patients will be screened to confirm study eligibility. The objectives of this study are to assess the safety and tolerability of OMP-313M32 in patients, to assess the maximum tolerated dose (MTD) of OMP-313M32, and to characterize the dose-limiting toxicity (DLT) of OMP-313M32 to determine the recommended Phase 2 dose of OMP-313M32; to characterize the pharmacokinetics of OMP-313M32; to characterize the immunogenic potential of OMP-313M32; Preliminary evaluation, pharmacokinetic (PD) markers.

As shown in Figure 2, dose escalation is performed to determine the maximum tolerated dose (MTD) or maximum administered dose (MAD) in the early phases of the trial. Dose levels of 0.3 mg/kg, 1.0 mg/kg, 3 mg/kg, 10 mg/kg, and 15 mg/kg of OMP-313M32 are administered by IV infusion once every two weeks. No dose escalation or reduction is permitted within a dose cohort. If no DLT is observed, 3 patients will be treated at each dose level. Once 1 in 3 patients experience a DLT, the dose level is escalated to 6 patients. Once 2 or more patients have experienced a DLT, no more patients will be dosed at this level, and 3 more patients will be dosed unless 6 patients are being treated at the above dose level. Patients are enrolled in the dose cohorts described above. Patients' DLT will be evaluated for 28 days (Days 1 through 29).

In the dose escalation phase of the study, patients will be enrolled at a dose level equal to the MTD or MAD of OMP-313M32. The expansion phase is designed to better characterize the safety and tolerability of OMP-313M32 and to preliminarily assess anti-tumor activity in patients with specific tumor types or subtypes. ing. Tumor types considered for evaluation in the dose escalation phase include head and neck cancer, esophageal/gastroesophageal cancer, gastric cancer, colorectal cancer, hepatocellular/liver cancer, cervical cancer, and lung cancer. , melanoma, Merkel cell carcinoma, renal cell/kidney cancer, bladder cancer, ovarian cancer, pancreatic cancer, endometrial cancer, and triple-negative breast cancer. As used herein, "lung cancer" includes non-small cell lung cancer (NSCLC) and small cell lung cancer. NSCLC can include NSCLC squamous and NSCLC adenocarcinoma. Colorectal cancer (CRC) can include high-frequency microsatellite-unstable CRC and microsatellite-stable CRC. Patients in the dose escalation phase had tumors that were resistant or refractory to treatment with PD-1 antagonists (e.g., anti-PD-1 antibodies) or PD-L1 antagonists (e.g., anti-PD-L1 antibodies). may Patients in the dose escalation phase have been previously treated with a PD-1 antagonist (e.g., anti-PD-1 antibody) or PD-L1 antagonist (e.g., anti-PD-L1 antibody) and are currently undergoing treatment or a subject with later progression of tumor growth (sometimes referred to as an anti-PD-1/PD-L1 progressor).

All patients enrolled in the dose escalation and escalation cohorts will have a biopsy prior to treatment, unless medically contraindicated, and at least one tumor biopsy after treatment begins. Biopsies are performed at baseline and approximately 2-3 weeks after the first dose of OMP-313M32. Additional biopsies may be collected at the discretion of the study director. Biopsy methods may include core needle biopsy, punch biopsy, forceps biopsy, or excisional/incisional biopsy. Expression of TIGIT, PVR, PVRL2, FOXP3, and other immune markers are assessed in FFPE tumor specimens. Expression levels of additional proteins and genes (eg, immune gene signatures) may be evaluated and correlated with clinical benefit.

Example 3
TIGIT protein expression assessed by IHC
To examine TIGIT protein expression, a TIGIT immunohistochemistry (IHC) assay was developed and optimized using a proprietary mouse anti-hTIGIT monoclonal antibody. Four micrometer FFPE sections were cut and mounted on positively-charged capillary gap slides. Tissues were dewaxed through four 5 minute changes of xylene followed by a graded alcohol series to distilled water. Steam heat-induced epitope recovery (SHIER) was performed with SHIER2 solution (citrate buffer pH 6.0-6.2) for 20 minutes in the upper chamber of a Black and Decker Steamer. Slides were pretreated with proteinase K enzyme (1:40 dilution) for 10 minutes and incubated in blocking solution for 15 minutes to reduce non-specific background staining. Slides were incubated overnight with 1 μg/ml anti-hTIGIT antibody. To block endogenous peroxidase activity, slides were incubated in hydrogen peroxide for 2.5 minutes three times and washed with PBS. Specific binding was detected using DAB and hematoxylin using the Polink-2 Plus Detection Kit designed for mouse antibodies. Positive staining is indicated by the presence of a brown chromogenic reaction product of horseradish peroxidase and DAB substrate. Blue nuclear staining is produced by hematoxylin counterstaining to assess cell and tissue morphology.

A panel of samples covering 17 different cancer types was evaluated for TIGIT staining in tumor cells, tumor-associated immune cells, and non-tumor/stromal immune cells by IHC assay (Table 2).

(Table 2)

Reactivity at the plasma membrane for tumor cells was assessed using the percent observed at differential intensities. Intensity scoring included 0=zero, 1+=low or weak, 2+=intermediate, and 3+=high or strong. The percentage recorded at each intensity level was reported from 0 to 100% in steps of 10 above 10% and 1 below 10%. Scoring data for TIGIT expression in tumor cells were evaluated using standard H-score and percentage scoring methods. H-scores were calculated using the percentage of cells with expression intensity (0, 1+, 2+, or 3+) on a three-point semiquantitative scale. Therefore, the scores range from 0-300. H-score = [(1+%) x 1] + [(2+%) x 2] + [(3+%) x 3]. Percentage scores were calculated by summing the percentages of intensity ≧1+, ≧2+, and ≧3+. Therefore, the scores range from 0-100. Percentage score > 1+ = (% of 1+) + (% of 2+) + (% of 3+). Percentage score > 2+ = (% of 2+) + (% of 3+). Percentage score > 3+ = (% of 3+).

Each sample is also tested for TIGIT reactivity in (1) immune cells in stromal/non-tumor areas and (2) immune cells in areas within or closely associated with tumors. analyzed. Immune cell reactivity was scored using a 0-3 abundance scale where 0 = no immune cell staining; 1 = little immune cell staining; 2 = moderate amount of immune cell staining; Abundant immune cell staining.

Table 3 summarizes the results of the IHC assay for TIGIT expression in tumor-associated immune cells.

(Table 3) TIGIT staining in tumor-associated immune cells

Overall, triple-negative breast cancer, T-cell lymphoma, head and neck cancer, and cervical cancer had the strongest staining of both stroma and tumor-associated immune cells. Consistent with this IHC data, analysis of 33 tumor types in The Cancer Genome Atlas (TGCA) by RNA-Seq showed a good correlation between TIGIT expression levels and T-cell markers. The overall incidence of TIGIT staining in the plasma membrane of tumor cells was low and weak in intensity. These results strongly suggest that TIGIT is expressed primarily in immune cells in tumors and the tumor microenvironment.

Example 4
Pharmacokinetic (PD) Biomarkers A multi-platform approach to examine the pharmacokinetic (PD) biomarkers of anti-TIGIT treatment in tumors and corresponding whole blood samples from tumor-bearing mice treated with the surrogate anti-TIGIT antibody 313R12. It was adopted. Tumor tissue and corresponding whole blood samples were analyzed for immune cell prevalence and function by microarray, immunochemistry (IHC), and flow cytometry.

Antibody 313R12 was administered to immunocompetent mice bearing CT26.WT colon tumors, 4T1 breast tumors, or B16F10 melanoma tumors. These tumors are believed to have varying levels of immunogenicity. CT26.WT tumors are highly immunogenic and enriched with cytotoxic T cells. 4T1 tumors are highly immunogenic and immunosuppressive. B16F10 tumors are hypoimmunogenic and immunologically silent. Mice were treated once weekly in each model.

To identify potential gene expression PD biomarkers that correlate with efficacy, RNA was isolated from tumor tissue and peripheral blood samples collected at study termination for each of the three models. Microarray analysis was performed using samples from the following groups: (1) 4T1 tumor-bearing mice treated with 0.1 mg/kg and 12.5 mg/kg 313R12, (2) 0.1 mg/kg and 12.5 mg/kg B16F10 tumor-bearing mice treated with kg 313R12, (3) CT26.WT tumor-bearing mice treated with 0.1 mg/kg, 0.5 mg/kg, 2.5 mg/kg, and 12.5 mg/kg 313R12, and (4) 3 animals from control mice. Global gene expression levels were profiled by microarray for treated and control tissues. Genetic alterations associated with immune cells were investigated, with an emphasis on T-cell, NK-cell, and B-cell markers. Changes in immune cell populations and cytokine secretion were monitored by flow cytometry, Luminex, and IHC.

In tumor samples, anti-TIGIT treatment promoted increased expression of genes associated with T cells, CD8 T cells, cytotoxic cells, Th1 cells, NK cells, Teff cells, and T cell activation markers. These gene expression changes were validated by quantitative real-time PCR (ie, CD3e, CD8a, Ncr1, Ifn-γ, Gzma, and CD266). Similar results were observed in gene expression changes in blood samples. These results suggest that PD biomarkers can be used in the clinic to study the on-target activity of the anti-TIGIT antibody OMP-313M32.

The examples and embodiments described herein are for illustrative purposes only and various modifications or alterations may be suggested to those skilled in the art in light of the examples and embodiments described herein and within the spirit of the present application. and is understood to be included within the scope.

All publications, patents, patent applications, internet sites, and accession numbers/database sequences, including both polynucleotide and polypeptide sequences, cited herein are each individual publications, patents, patent applications. , internet sites, or accession numbers/database sequences are hereby incorporated by reference in their entirety for all purposes to the same extent as if they were specifically and individually indicated to be incorporated by reference.

The sequences disclosed in this application are:

Claims (27)

A medicament for use in a method of inhibiting cancer in a human patient with cancer comprising a therapeutically effective amount of a full-length antibody that specifically binds to the extracellular domain of human TIGIT, comprising:
said method comprising administering said therapeutically effective amount of a full-length antibody to said human patient;
The cancer is head and neck cancer, cervical cancer, gastric cancer, ovarian cancer, melanoma, sarcoma, lung cancer, gastroesophageal cancer, leukemia, lymphoma, anal cancer, pancreatic cancer, hepatocellular carcinoma, Liver cancer, renal cell carcinoma, kidney cancer, bladder cancer, high-frequency microsatellite-unstable colorectal cancer, microsatellite-stable colorectal cancer, triple-negative breast cancer, Merkel cell carcinoma, endometrial cancer cancer, or esophageal cancer, and the full-length antibody that binds to human TIGIT is capable of mediating antibody-dependent cellular cytotoxicity (ADCC) and/or complement-dependent cytotoxicity (CDC), and SEQ ID a heavy chain variable region comprising NO:10 and a light chain variable region comprising SEQ ID NO:11 ;
Medicine. said human patient has been previously treated with a human PD-1 antagonist or human PD-L1 antagonist and during or during treatment with said human PD-1 antagonist or said human PD-L1 antagonist 2. The medicament of claim 1, followed by tumor growth, progression or recurrence. 2. The medicament of claim 1, wherein said antibody that binds to human TIGIT comprises the heavy chain amino acid sequence of SEQ ID NO:13 and the light chain amino acid sequence of SEQ ID NO:15. wherein said antibody that binds to human TIGIT is a heavy chain variable region encoded by a plasmid deposited with ATCC under designation number PTA-122346 and a light chain variable region encoded by a plasmid deposited with ATCC under designation number PTA-122347; 2. A medicament according to claim 1, comprising a region. The method further comprises administering to the patient an additional human therapeutic agent, wherein the additional human therapeutic agent is administered prior to, concurrently with, or after administration of the antibody that binds to the human TIGIT; 2. The medicament of claim 1, which is administered. The method administers to the patient at least one additional therapeutic agent selected from the group consisting of nivolumab, pembrolizumab, atezolizumab, avelumab, ipilimumab, bevacizumab, ramucirumab, trastuzumab, pertuzumab, panitumumab, cetuximab, semiplimab, and durvalumab. 2. The medicament of claim 1, further comprising a step, wherein said additional therapeutic agent is administered prior to, concurrently with, or after administration of said antibody that binds to human TIGIT. 7. The medicament of claim 6 , wherein said additional therapeutic agent is selected from the group consisting of nivolumab, pembrolizumab and ipilimumab. 2. The medicament of claim 1, wherein said antibody that binds to human TIGIT is administered intravenously to said human patient. 2. The medicament of claim 1, wherein said antibody that binds to human TIGIT is administered once a week, once every two weeks, once every three weeks, or once every four weeks. said antibody that binds to human TIGIT comprises the heavy chain amino acid sequence of SEQ ID NO:13 and the light chain amino acid sequence of SEQ ID NO:15;
an antibody that binds to said human TIGIT is administered intravenously to said human patient;
wherein said antibody that binds to human TIGIT is administered once weekly, once every two weeks, once every three weeks, or once every four weeks, and said method administers at least one additional human therapeutic to said patient; administering, wherein said additional human therapeutic is administered prior to, concurrently with, or after administration of said antibody that binds to said human TIGIT;
The medicament according to claim 1. A medicament for use in a method of inhibiting cancer in a human patient with cancer comprising a therapeutically effective amount of a full-length antibody that specifically binds to the extracellular domain of human TIGIT, comprising:
said method comprising administering said therapeutically effective amount of a full-length antibody to said human patient;
said human patient has been previously treated with a human PD-1 antagonist or a human PD-L1 antagonist and during or during treatment with said human PD-1 antagonist or said human PD-L1 antagonist is followed by tumor growth, progression, or recurrence, and full-length antibodies that bind to said human TIGIT can mediate antibody-dependent cellular cytotoxicity (ADCC) and/or complement-dependent cytotoxicity (CDC); and a heavy chain variable region comprising SEQ ID NO:10 and a light chain variable region comprising SEQ ID NO:11 ,
Medicine. 12. The medicament of claim 11, wherein said antibody that binds to human TIGIT comprises the heavy chain amino acid sequence of SEQ ID NO:13 and the light chain amino acid sequence of SEQ ID NO:15. The method further comprises administering to the patient an additional human therapeutic agent, wherein the additional human therapeutic agent is administered prior to, concurrently with, or after administration of the antibody that binds to the human TIGIT; 12. A medicament according to claim 11 , which is administered. The method administers to the patient at least one additional therapeutic agent selected from the group consisting of nivolumab, pembrolizumab, atezolizumab, avelumab, ipilimumab, bevacizumab, ramucirumab, trastuzumab, pertuzumab, panitumumab, cetuximab, semiplimab, and durvalumab. 14. The medicament of claim 13 , further comprising a step, wherein said additional human therapeutic is administered prior to, concurrently with, or after administration of said antibody that binds to human TIGIT. 15. The medicament of claim 14 , wherein said additional therapeutic agent is selected from the group consisting of nivolumab, pembrolizumab and ipilimumab. 12. The medicament of claim 11 , wherein said antibody that binds to human TIGIT is administered intravenously to said human patient. 12. The medicament of claim 11 , wherein said antibody that binds to human TIGIT is administered once a week, once every two weeks, once every three weeks, or once every four weeks. The cancer is head and neck cancer, cervical cancer, gastric cancer, ovarian cancer, melanoma, sarcoma, lung cancer, gastroesophageal cancer, leukemia, lymphoma, anal cancer, pancreatic cancer, hepatocellular carcinoma, Liver cancer, renal cell carcinoma, kidney cancer, bladder cancer, high-frequency microsatellite-unstable colorectal cancer, microsatellite-stable colorectal cancer, triple-negative breast cancer, Merkel cell carcinoma, endometrial cancer 12. The pharmaceutical according to claim 11 , which is cancer or esophageal cancer. said antibody that binds to human TIGIT comprises the heavy chain amino acid sequence of SEQ ID NO:13 and the light chain amino acid sequence of SEQ ID NO:15;
an antibody that binds to said human TIGIT is administered intravenously to said human patient;
wherein said antibody that binds to human TIGIT is administered once weekly, once every two weeks, once every three weeks, or once every four weeks, and said method administers at least one additional human therapeutic to said patient; 12. The medicament of claim 11 , further comprising administering, wherein said additional human therapeutic is administered prior to, concurrently with, or after administration of said antibody that binds to said human TIGIT. A medicament for use in a method of inhibiting cancer in a human patient with cancer comprising a therapeutically effective amount of a full-length antibody that specifically binds to the extracellular domain of human TIGIT, comprising:
said method comprising administering said therapeutically effective amount of a full-length antibody to said human patient;
the cancer expresses poliovirus receptor (PVR) and/or poliovirus receptor-related 2 (PVRL2), and the full-length antibody that binds to the human TIGIT causes antibody-dependent cellular cytotoxicity (ADCC) and/or capable of mediating complement dependent cytotoxicity (CDC) and comprising a heavy chain variable region comprising SEQ ID NO:10 and a light chain variable region comprising SEQ ID NO:11 ;
Medicine. 21. The medicament of claim 20, wherein said antibody that binds to human TIGIT comprises the heavy chain amino acid sequence of SEQ ID NO:13 and the light chain amino acid sequence of SEQ ID NO:15. The method further comprises administering to the patient an additional human therapeutic agent, wherein the additional human therapeutic agent is administered prior to, concurrently with, or after administration of the antibody that binds to the human TIGIT; 21. A medicament according to claim 20 , which is administered. The method administers to the patient at least one additional therapeutic agent selected from the group consisting of nivolumab, pembrolizumab, atezolizumab, avelumab, ipilimumab, bevacizumab, ramucirumab, trastuzumab, pertuzumab, panitumumab, cetuximab, semiplimab, and durvalumab. 23. The medicament of claim 22 , further comprising a step, wherein said additional human therapeutic is administered prior to, concurrently with, or after administration of said antibody that binds to human TIGIT. 23. The medicament of Claim 22 , wherein said additional therapeutic agent is selected from the group consisting of nivolumab, pembrolizumab, and ipilimumab. said antibody that binds to human TIGIT is administered intravenously to said human patient, and said antibody that binds to human TIGIT is administered once weekly, once every two weeks, once every three weeks, or once every four weeks 21. A medicament according to claim 20 , which is administered. The cancer is head and neck cancer, cervical cancer, gastric cancer, ovarian cancer, melanoma, sarcoma, lung cancer, gastroesophageal cancer, leukemia, lymphoma, anal cancer, pancreatic cancer, hepatocellular carcinoma, Liver cancer, renal cell carcinoma, kidney cancer, bladder cancer, high-frequency microsatellite-unstable colorectal cancer, microsatellite-stable colorectal cancer, triple-negative breast cancer, Merkel cell carcinoma, endometrial cancer 21. The pharmaceutical according to claim 20 , which is cancer or esophageal cancer. said antibody that binds to human TIGIT comprises the heavy chain amino acid sequence of SEQ ID NO:13 and the light chain amino acid sequence of SEQ ID NO:15;
an antibody that binds to said human TIGIT is administered intravenously to said human patient;
wherein said antibody that binds to human TIGIT is administered once weekly, once every two weeks, once every three weeks, or once every four weeks, and said method administers at least one additional human therapeutic to said patient; 21. The medicament of claim 20 , further comprising administering, wherein said additional human therapeutic is administered prior to, concurrently with, or after administration of said antibody that binds to said human TIGIT.

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